Hydroxyapatite coating affects the Wnt signaling pathway during peri-implant healing in vivo

Owing to its bio- and osteoconductivity, hydroxyapatite (HA) is a widely used implant material, but its osteogenic properties are only partly evaluated in vitro and in vivo. The present study focused on bone healing adjacent to HA-coated titanium (Ti) implants, with or without incorporated lithium ions (Li⁺). Special attention was given to the Wnt signaling pathway. The implants were inserted into rat tibia for 7 or 28 days and analyzed ex vivo, mainly by histomorphometry and quantitative real-time polymerase chain reaction (qPCR). HA-coated implants showed, irrespective of Li⁺ content, bone–implant contact (BIC) and removal torque values significantly higher than those of reference Ti. Further, the expression of OCN, CTSK, COL1A1, LRP5/6 and WISP1 was significantly higher in implant-adherent cells of HA-coated implants, with or without Li⁺. Significantly higher β-catenin expression and significantly lower COL2A1 expression were observed in peri-implant bone cells from HA with 14 ng cm⁻² released Li⁺. Interestingly, Ti implants showed a significantly larger bone area (BA) in the threads than HA with 39 ng cm⁻² released Li⁺, but had a lower BIC than any HA-coated implant. This study shows that HA, with or without Li⁺, is a strong activator of the Wnt signaling pathway, and may to some degree explain its high bone induction capacity.     

Ultraviolet light-mediated photofunctionalization of titanium to promote human mesenchymal stem cell migration,attachment, proliferation and differentiation

Improving the osteoconductive potential of titanium implants has been of continuing interest in the fields of dentistry and orthopedic surgery. This study determined the bioactivity of ultraviolet (UV) light-treated titanium. Human mesenchymal stem cells (MSCs) were cultured on acid-etched microtopographical titanium surfaces with and without 48 h pretreatment with UVA (peak wavelength of 360 nm) or UVC (peak wavelength of 250 nm). The number of cells that migrated to the UVC-treated surface during the first 3 h of incubation was eight times higher than those that migrated to the untreated surface. After 24 h of incubation, the number of cells attached to the UVC-treated surface was over three times more than those attached to the untreated surface. On the UVC-treated surface, the cellular spread was expedited with an extensive and intensive expression of the focal adhesion protein vinculin. The cells on the UVC-treated surface exhibited a threefold higher bromodeoxyuridine incorporation, a doubling of the alkaline phosphatase-positive area and the up-regulated expression of bone-related genes, indicating the accelerated proliferation and differentiation. The UVC-treated surface did not adversely affect the viability of the cells. These biological effects were not seen after UVA treatment, despite the generation of superhydrophilicity. Thus, we discovered a novel photofunctionalization of titanium dioxide that substantially enhances its bioactivity in human MSCs. Further studies are required to investigate the universal effectiveness of this surface modification for different titanium-containing materials, with varying chemistries and textures, as well as to understand its significance in enhancing in vivo osteoconductivity.     

Comparison of two carbonated apatite ceramics in vivo

Carbonated apatite ceramics, with a composition similar to that of bone mineral, are potentially interesting synthetic bone graft substitutes. In the present study, two porous carbonated apatite ceramics were developed, characterized and tested for their bone repair capacity and osteoinductive potential in a goat model. Although the two ceramics were prepared from a similar starting powder, their physico-chemical and structural characteristics differed as a consequence of different preparation methods. Both ceramics had an open and interconnected porous structure with a porosity of about 80%. The morphology of the surface of CA-A and CA-B at the submicron level differed significantly: CA-A consisted of irregular grains with a size of 5–20 μm, with 1–10 μm large micropores among the grains, whereas CA-B surface consisted of much smaller and regular shaped grains (0.05–0.5 μm), with most micropores smaller than 1 μm. The specific surface area of CA-B was about 10 times larger than that of CA-A due to its significantly smaller grain size. CA-A and CA-B ceramics contained 3 and 5 wt.% of B-type carbonated apatite, respectively. Although neither ceramic succeeded in completely bridging the 17 mm iliac wing defect with new bone after 12 weeks of implantation, CA-A showed significantly more bone formation in the pores of the implant than CA-B. The total area percentage of new bone in the total defect area was 12.7 ± 1.81 and 5.51 ± 1.37 (mean ± SEM) for CA-A and CA-B, respectively. Intramuscular implantation of the ceramics led to ectopic bone formation by CA-A in all three implanted specimens, in contrast to CA-B, where no new bone was observed in any of the 11 animals. CA-A showed a more pronounced degradation than CA-B both in vitro and in vivo at both implantation sites, which was unexpected based on the physico-chemical and structural properties of the two ceramics. Both physico-chemical and structural properties of the ceramics could, dependently or independently, have affected their in vivo behaviour, emphasizing the importance to control individual parameters for successful bone repair.     

Multi-functional P(3HB) microsphere/45S5 Bioglass®-based composite scaffolds for bone tissue engineering

Novel multi-functional P(3HB) microsphere/45S5 Bioglass®-based composite scaffolds exhibiting potential for drug delivery were developed for bone tissue engineering. 45S5 Bioglass®-based glass–ceramic scaffolds of high interconnected porosity produced using the foam-replication technique were coated with biodegradable microspheres (size < 2 μm) made from poly(3-hydroxybutyrate), P(3HB), produced using Bacillus cereus SPV. A solid-in-oil-in-water emulsion solvent extraction/evaporation technique was used to produce these P(3HB) microspheres. A simple slurry-dipping method, using a 1 wt.% suspension of P(3HB) microspheres in water, dispersed by an ultrasonic bath, was used to coat the scaffold, producing a uniform microsphere coating throughout the three-dimensional scaffold structure. Compressive strength tests confirmed that the microsphere coating slightly enhanced the scaffold mechanical strength. It was also confirmed that the microsphere coating did not inhibit the bioactivity of the scaffold when immersed in simulated body fluid (SBF) for up to 4 weeks. The hydroxyapatite (HA) growth rate on P(3HB) microsphere-coated 45S5 Bioglass® composite scaffolds was very similar to that on the uncoated control sample, qualitatively indicating similar bioactivity. However, the surface topography of the HA surface layer was affected as shown by results obtained from white light interferometry. The roughness of the surface was much higher for the P(3HB) microsphere-coated scaffolds than for the uncoated samples, after 7 days in SBF. This feature would facilitate cell attachment and proliferation. Finally, gentamycin was successfully encapsulated into the P(3HB) microspheres to demonstrate the drug delivery capability of the scaffolds. Gentamycin release kinetics was determined using liquid chromatography–mass spectrometry. The release of the drug from the coated composite scaffolds was slow and controlled when compared to the observed fast and relatively uncontrolled drug release from the bone scaffold (without microsphere coating). Thus, this unique multifunctional bioactive composite scaffold has the potential to enhance cell attachment and to provide controlled delivery of relevant drugs for bone tissue engineering.     

Enhanced bone regeneration in rat calvarial defects implanted with surface-modified and BMP-loaded bioactive glass (13-93) scaffolds

The repair of large bone defects, such as segmental defects in the long bones of the limbs, is a challenging clinical problem. Our recent work has shown the ability to create porous scaffolds of silicate 13-93 bioactive glass by robocasting which have compressive strengths comparable to human cortical bone. The objective of this study was to evaluate the capacity of those strong porous scaffolds with a grid-like microstructure (porosity = 50%; filament width = 330 μm; pore width = 300 μm) to regenerate bone in a rat calvarial defect model. Six weeks post-implantation, the amount of new bone formed within the implants was evaluated using histomorphometric analysis. The amount of new bone formed in implants composed of the as-fabricated scaffolds was 32% of the available pore space (area). Pretreating the as-fabricated scaffolds in an aqueous phosphate solution for 1, 3 and 6 days to convert a surface layer to hydroxyapatite prior to implantation enhanced new bone formation to 46%, 57% and 45%, respectively. New bone formation in scaffolds pretreated for 1, 3 and 6 days and loaded with bone morphogenetic protein-2 (BMP-2) (1 μg per defect) was 65%, 61% and 64%, respectively. The results show that converting a surface layer of the glass to hydroxyapatite or loading the surface-treated scaffolds with BMP-2 can significantly improve the capacity of 13-93 bioactive glass scaffolds to regenerate bone in an osseous defect. Based on their mechanical properties evaluated previously and their capacity to regenerate bone found in this study, these 13-93 bioactive glass scaffolds, pretreated or loaded with BMP-2, are promising in structural bone repair.     

Comparison of enhancement of bone ingrowth into hydroxyapatite ceramics with highly and poorly interconnected pores by electrical polarization

The effects of electrical polarization of porous hydroxyapatite ceramics with different structures on bone ingrowth were compared. Two types of cylindrical porous hydroxyapatite ceramics with high and low interpore connection (hydroxyapatite-H and hydroxyapatite-L, respectively) were utilized in this study. Hydroxyapatite-H or hydroxyapatite-L with and without electrical polarization was implanted into the right or left femoral condyle of rabbits (n = 10 in each group) and histological examination was performed 3 and 6 weeks after operation. Each cross-section was divided into three regions, outer, middle and inner region, and the percentage of total newly formed bone area/total area of each region (% bone area) was calculated. Bone ingrowth throughout the region of implant was significantly larger in the hydroxyapatite-H group than in the hydroxyapatite-L group. Electrical polarization was effective in enhancing bone ingrowth through all the pores of hydroxyapatite-H implant, however, this advantage was not apparent in the hydroxyapatite-L implant. It is suggested that enhanced bone ingrowth into hydroxyapatite porous bodies due to electrical polarization may be a cooperative interaction between the osteoconductivity of hydroxyapatite porous bodies and enhanced osteogenic cell activity induced by large charges stored on the pore surfaces.     

Bone attachment to glass-fibre-reinforced composite implant with porous surface

A method has recently been developed for producing fibre-reinforced composites (FRC) with porous surfaces, intended for use as load-bearing orthopaedic implants. This study focuses on evaluation of the bone-bonding behaviour of FRC implants. Three types of cylindrical implants, i.e. FRC implants with a porous surface, solid polymethyl methacrylate (PMMA) implants and titanium (Ti) implants, were inserted in a transverse direction into the intercondular trabeculous bone area of distal femurs and proximal tibias of New Zealand White rabbits. Animals were sacrificed at 3, 6 and 12 weeks post operation, and push-out tests (n = 5–6 per implant type per time point) were then carried out. At 12 weeks the shear force at the porous FRC–bone interface was significantly higher (283.3 ± 55.3 N) than the shear force at interfaces of solid PMMA/bone (14.4 ± 11.0 N; p < 0.001) and Ti/bone (130.6 ± 22.2 N; p = 0.001). Histological observation revealed new bone growth into the porous surface structure of FRC implants. Solid PMMA and Ti implants were encapsulated mostly with fibrous connective tissue. Finite element analysis (FEA) revealed that porous FRC implants had mechanical properties which could be tailored to smooth the shear stress distribution at the bone–implant interface and reduce the stress-shielding effect.     

Bone regeneration in strong porous bioactive glass (13-93) scaffolds with an oriented microstructure implanted in rat calvarial defects

There is a need for synthetic bone graft substitutes to repair large bone defects resulting from trauma, malignancy and congenital diseases. Bioactive glass has attractive properties as a scaffold material but factors that influence its ability to regenerate bone in vivo are not well understood. In the present work, the ability of strong porous scaffolds of 13-93 bioactive glass with an oriented microstructure to regenerate bone was evaluated in vivo using a rat calvarial defect model. Scaffolds with an oriented microstructure of columnar pores (porosity = 50%; pore diameter = 50?150 μm) showed mostly osteoconductive bone regeneration, and new bone formation, normalized to the available pore area (volume) of the scaffolds, increased from 37% at 12 weeks to 55% at 24 weeks. Scaffolds of the same glass with a trabecular microstructure (porosity = 80%; pore width = 100?500 μm), used as the positive control, showed bone regeneration in the pores of 25% and 46% at 12 and 24 weeks, respectively. The brittle mechanical response of the as-fabricated scaffolds changed markedly to an elastoplastic response in vivo at both implantation times. These results indicate that both groups of 13-93 bioactive glass scaffolds could potentially be used to repair large bone defects, but scaffolds with the oriented microstructure could also be considered for the repair of loaded bone.     

Loss of mechanical properties in vivo and bone–implant interface strength of AZ31B magnesium alloy screws with Si-containing coating

In this study the loss of mechanical properties and the interface strength of coated AZ31B magnesium alloy (a magnesium–aluminum alloy) screws with surrounding host tissues were investigated and compared with non-coated AZ31B, degradable polymer and biostable titanium alloy screws in a rabbit animal model after 1, 4, 12 and 21 weeks of implantation. The interface strength was evaluated in terms of the extraction torque required to back out the screws. The loss of mechanical properties over time was indicated by one-point bending load loss of the screws after these were extracted at different times. AZ31B samples with a silicon-containing coating had a decreased degradation rate and improved biological properties. The extraction torque of Ti6Al4V, poly-l-lactide (PLLA) and coated AZ31B increased significantly from 1 week to 4 weeks post-implantation, indicating a rapid osteosynthesis process over 3 weeks. The extraction torque of coated AZ31B increased with implantation time, and was higher than that of PLLA after 4 weeks of implantation, equalling that of Ti6Al4V at 12 weeks and was higher at 21 weeks. The bending loads of non-coated AZ31B and PLLA screws degraded sharply after implantation, and that of coated AZ31B degraded more slowly. The biodegradation mechanism, the coating to control the degradation rate and the bioactivity of magnesium alloys influencing the mechanical properties loss over time and bone–implant interface strength are discussed in this study and it is concluded that a suitable degradation rate will result in an improvement in the mechanical performance of magnesium alloys, making them more suitable for clinical application.     

Peri- and intra-implant bone response to microporous Ti coatings with surface modification

Bone growth on and into implants exhibiting substantial surface porosity is a promising strategy in order to improve the long-term stable fixation of bone implants. However, the reliability in clinical applications remains a point of discussion. Most attention has been dedicated to the role of macroporosity, leading to the general consensus of a minimal pore size of 50–100 μm in order to allow bone ingrowth. In this in vivo study, we assessed the feasibility of early bone ingrowth into a predominantly microporous Ti coating with an average thickness of 150 μm and the hypothesis of improving the bone response through surface modification of the porous coating. Implants were placed in the cortical bone of rabbit tibiae for periods of 2 and 4 weeks and evaluated histologically and histomorphometrically using light microscopy and scanning electron microscopy. Bone with osteocytes encased in the mineralized matrix was found throughout the porous Ti coating up to the coating/substrate interface, highlighting that osseointegration of microporosities (<10 μm) was achievable. The bone trabeculae interweaved with the pore struts, establishing a large contact area which might enable an improved load transfer and stronger implant/bone interface. Furthermore, there was a clear interconnection with the surrounding cortical bone, suggesting that mechanical interlocking of the coating in the host bone in the long term is possible. When surface modifications inside the porous structure further reduced the interconnective pore size to the submicrometer level, bone ingrowth was impaired. On the other hand, application of a sol–gel-derived bioactive glass–ceramic coating without altering the pore characteristics was found to significantly improve bone regeneration around the coating, while still supporting bone ingrowth.     

Poly(l-glutamic acid)/chitosan polyelectrolyte complex porous microspheres as cell microcarriers for cartilage regeneration

In this study a novel kind of porous poly(l-glutamic acid) (PLGA)/chitosan polyelectrolyte complex (PEC) microsphere was developed through electrostatic interaction between PLGA and chitosan. By adjusting the formula parameters chitosan microspheres with an average pore size of 47.5 ± 5.4 μm were first developed at a concentration of 2 wt.% and freeze temperature of −20 °C. For self-assembly of the PEC microspheres porous chitosan microspheres were then incubated in PLGA solution at 37 °C. Due to electrostatic interaction a large amount of PLGA (110.3 μg mg⁻¹) was homogeneously absorbed within the chitosan microspheres. The developed PEC microspheres retained their original size, pore diameters and interconnected porous structure. Fourier transform infrared spectroscopy, thermal gravimetric analysis and zeta potential analysis revealed that the PEC microspheres were successfully prepared through electrostatic interaction. Compared with microspheres fabricated from chitosan, the porous PEC microspheres were shown to efficiently promote chondrocyte attachment and proliferation. After injection subcutaneously for 8 weeks PEC microspheres loaded with chondrocytes were found to produce significant more cartilaginous matrix than chitosan microspheres. These results indicate that these novel fabricated porous PLGA/chitosan PEC microspheres could be used as injectable cell carriers for cartilage tissue engineering.     

The effect of UV-photofunctionalization on the time-related bioactivity of titanium and chromium–cobalt alloys

This study examined the possible changes in the bioactivity of titanium surfaces during their aging and investigated the effect of ultraviolet (UV) light treatment during the age-related change of titanium bioactivity. Rat bone marrow-derived osteoblastic cells were cultured on new titanium disks (immediately after either acid-etching, machining, or sandblasting), 4-week-old disks (stored after processing for 4 weeks in dark ambient conditions), and 4-week-old disks treated with UVA (peak wavelength of 365 nm) or UVC (peak wavelength of 250 nm). During incubation for 24 h, only 50% of the cells were attached to the 4-week-old surfaces as compared to the new surface. UVC treatment of the aged surface increased its cell attachment capacity to a level 50% higher than the new surfaces, whereas UVA treatment had no effect. Proliferation, alkaline phosphatase activity, and mineralization of cells were substantially lower on the 4-week-old surfaces than on the new surfaces, while they were higher on the UVC-treated 4-week-old surfaces as compared to the new surfaces. The age-related impaired bioactivity was found on all titanium topographies as well as on a chromium–cobalt alloy, and was associated with an increased percentage of surface carbon. Although both UVA and UVC treatment converted the 4-week-old titanium surfaces from hydrophobic to superhydrophilic, only UVC treatment effectively reduced the surface carbon to a level equivalent to the new surface. Thus, this study uncovered a time-dependent biological degradation of titanium and chromium–cobalt alloy, and its restoration enabled by UVC phototreatment, which surmounts the innate bioactivity of new surfaces, which is more closely linked to hydrocarbon removal than the induced superhydrophilicity.     

Repairing a critical-sized bone defect with highly porous modified and unmodified baghdadite scaffolds

This is the first reported study to prepare highly porous baghdadite (Ca₃ZrSi₂O₉) scaffolds with and without surface modification and investigate their ability to repair critical-sized bone defects in a rabbit radius under normal load. The modification was carried out to improve the mechanical properties of the baghdadite scaffolds (particularly to address their brittleness) by coating their surfaces with a thin layer (～400 nm) of polycaprolactone (PCL)/bioactive glass nanoparticles (nBGs). The β-tricalcium phosphate/hydroxyapatite (TCP/HA) scaffolds with and without modification were used as the control groups. All of the tested scaffolds had an open and interconnected porous structure with a porosity of ～85% and average pore size of 500 μm. The scaffolds (six per scaffold type and size of 4 mm × 4 mm × 15 mm) were implanted (press-fit) into the rabbit radial segmental defects for 12 weeks. Micro-computed tomography and histological evaluations were used to determine bone ingrowth, bone quality, and implant integration after 12 weeks of healing. Extensive new bone formation with complete bridging of the radial defect was evident with the baghdadite scaffolds (modified/unmodified) at the periphery and in close proximity to the ceramics within the pores, in contrast to TCP/HA scaffolds (modified/unmodified), where bone tended to grow between the ulna adjacent to the implant edge. Although the modification of the baghdadite scaffolds significantly improved their mechanical properties, it did not show any significant effect on in vivo bone formation. Our findings suggest that baghdadite scaffolds with and without modification can serve as a potential material to repair critical sized bone defects.     

Cellular activity of bioactive nanograined/ultrafine-grained materials

Our recent electron microscopy study on biomimetic nanostructured coatings on nanograined/ultrafine-grained (NG/UFG) substrates [Mater Sci Eng C 2009;29:2417–27] indicated that electrocrystallized nanohydroxyapatite (nHA) on phase-reversion-induced NG/UFG substrates exhibited a vein-type interconnected and fibrillar structure that closely mimicked the hierarchical structure of bone. The fibrillar structure on NG/UFG substrate is expected to be more favorable for cellular response than a planar surface. In contrast, hydroxyapatite (HA) coating on coarse-grained (CG) substrate more closely resembled a film rather than a fibrillar structure. Inspired by the differences in the structure of HA coating, we describe here the cell–substrate interactions of pre-osteoblasts (MC 3T3-E1) on bioactive NG/UFG and CG austenitic stainless steel substrates. NG/UFG austenitic stainless steel was obtained by a novel controlled phase-reversion annealing of cold-deformed austenite. This example provides an illustration of how a combination of cellular and molecular biology, materials science and engineering can advance our understanding of cell–substrate interactions. Interestingly, the cellular response of nanohydroxyapatite (nHA)-coated NG/UFG substrate demonstrated superior cytocompatibility, improved initial cell attachment, higher viability and proliferation, and well-spread morphology in relation to HA-coated CG substrate and their respective uncoated (bare) counterparts as implied by fluorescence and electron microscopy and MTT assay. Similar conclusions were derived from an immunofluorescence study that involved examination of the expression levels of vinculin focal adhesion contacts associated with dense actin stress fibers and fibronectin, protein analysis through protein bands in SDS–PAGE, and quantitative total protein assay. The enhancement of cellular response followed the sequence: nHA-coated NG/UFG > nHA-coated CG > NG/UFG > CG substrates. The outcomes of the study are expected to counter the challenges associated with the engineering of nanostructured surfaces with specific physical and surface properties for medical devices with significantly improved cellular response.     

Reconstruction of goat femur segmental defects using triphasic ceramic-coated hydroxyapatite in combination with autologous cells and platelet-rich plasma

Segmental bone defects resulting from trauma or pathology represent a common and significant clinical problem. In this study, a triphasic ceramic (calcium silicate, hydroxyapatite and tricalcium phosphate)-coated hydroxyapatite (HASi) having the benefits of both HA (osteointegration, osteoconduction) and silica (degradation) was used as a bone substitute for the repair of segmental defect (2 cm) created in a goat femur model. Three experimental goat femur implant groups – (a) bare HASi, (b) osteogenic-induced goat bone marrow-derived mesenchymal stem cells cultured HASi (HASi + C) and (c) osteogenic-induced goat bone marrow-derived mesenchymal stem cells cultured HASi + platelet-rich plasma (HASi + CP) – were designed and efficacy performance in the healing of the defect was evaluated. In all the groups, the material united with host bone without any inflammation and an osseous callus formed around the implant. This reflects the osteoconductivity of HASi where the cells have migrated from the cut ends of host bone. The most observable difference between the groups appeared in the mid region of the defect. In bare HASi groups, numerous osteoblast-like cells could be seen together with a portion of material. However, in HASi + C and HASi + CP, about 60–70% of that area was occupied by woven bone, in line with material degradation. The interconnected porous nature (50–500 μm), together with the chemical composition of the HASi, facilitated the degradation of HASi, thereby opening up void spaces for cellular ingrowth and bone regeneration. The combination of HASi with cells and PRP was an added advantage that could promote the expression of many osteoinductive proteins, leading to faster bone regeneration and material degradation. Based on these results, we conclude that bare HASi can aid in bone regeneration but, with the combination of cells and PRP, the sequence of healing events are much faster in large segmental bone defects in weight-bearing areas in goats.     

Geometric analysis of porous bone substitutes using micro-computed tomography and fuzzy distance transform

There is increased interest in resorbable bone substitutes for skeletal reconstruction. Important geometric design measures of bone substitute include pore size, interconnection size, porosity, permeability and surface area of the substitute. In this study, four substitute groups with variable geometric features but constant porosity were scanned using micro-computed tomography (μCT) and their geometric measures were determined using an advanced image-processing algorithm based on fuzzy distance transform and new pore size definition. The substitutes were produced using the calcium phosphate emulsion method. The geometric analysis revealed that the reproducibility of the emulsion method was high, within 5%. The average porosity of the four groups was 52.3 ± 1.5. The pore diameter of the four bone substitute groups was measured to be 170 ± 1.7, 217 ± 5.2, 416 ± 19, and 972 ± 11 μm. Despite this significant change in pore size, the interconnection size only increased slightly with an increase of pore size. The specific surface decreased with increasing pore size. The permeability increased with the pore size and was inversely proportional to the specific surface. The combination of μCT and the fuzzy image-processing tool enables accurate geometric analysis, even if pore size and image resolution are in the same range, such as in the case of the smallest pore size. Moreover, it is an exciting tool to understand the structure of the substitute with the hope of designing better bone substitutes.     

In vitro and in vivo evaluation of the surface bioactivity of a calcium phosphate coated magnesium alloy

Magnesium has shown potential application as a bio-absorbable biomaterial, such as for bone screws and plates. In order to improve the surface bioactivity, a calcium phosphate was coated on a magnesium alloy by a phosphating process (Ca–P coating). The surface characterization showed that a porous and netlike CaHPO₄·2H₂O layer with small amounts of Mg²⁺ and Zn²⁺ was formed on the surface of the Mg alloy. Cells L929 showed significantly good adherence and significantly high growth rate and proliferation characteristics on the Ca–P coated magnesium alloy (p < 0.05) in in-vitro cell experiments, demonstrating that the surface cytocompatibility of magnesium was significantly improved by the Ca–P coating. In vivo implantations of the Ca–P coated and the naked alloy rods were carried out to investigate the bone response at the early stage. Both routine pathological examination and immunohistochemical analysis demonstrated that the Ca–P coating provided magnesium with a significantly good surface bioactivity (p < 0.05) and promoted early bone growth at the implant/bone interface. It was suggested that the Ca–P coating might be an effective method to improve the surface bioactivity of magnesium alloy.     

Unique microstructural design of ceramic scaffolds for bone regeneration under load

During the past two decades, research on ceramic scaffolds for bone regeneration has progressed rapidly; however, currently available porous scaffolds remain unsuitable for load-bearing applications. The key to success is to apply microstructural design strategies to develop ceramic scaffolds with mechanical properties approaching those of bone. Here we report on the development of a unique microstructurally designed ceramic scaffold, strontium–hardystonite–gahnite (Sr–HT–gahnite), with 85% porosity, 500 μm pore size, a competitive compressive strength of 4.1 ± 0.3 MPa and a compressive modulus of 170 ± 20 MPa. The in vitro biocompatibility of the scaffolds was studied using primary human bone-derived cells. The ability of Sr–HT–gahnite scaffolds to repair critical-sized bone defects was also investigated in a rabbit radius under normal load, with β-tricalcium phosphate/hydroxyapatite scaffolds used in the control group. Studies with primary human osteoblast cultures confirmed the bioactivity of these scaffolds, and regeneration of rabbit radial critical defects demonstrated that this material induces new bone defect bridging, with clear evidence of regeneration of original radial architecture and bone marrow environment.     

Increased bone remodelling around titanium implants coated with chondroitin sulfate in ovariectomized rats

Coating titanium implants with artificial extracellular matrices based on collagen and chondroitin sulfate (CS) has been shown to enhance bone remodelling and de novo bone formation in vivo. The aim of this study was to evaluate the effect of estrogen deficiency and hormone replacement therapy (HRT) on the osseointegration of CS-modified Ti implants. 30 adult female, ovariectomized Wistar rats were fed either with an ethinyl-estradiol-rich diet (E) to simulate a clinical relevant HRT or with a genistein-rich diet (G) to test an alternative therapy based on nutritionally relevant phytoestrogens. Controls (C) received an estrogen-free diet. Uncoated titanium pins (Ti) or pins coated with type-I collagen and CS (Ti/CS) were inserted 8 weeks after ovarectomy into the tibia. Specimens were retrieved 28 days after implantation. Both the amount of newly formed bone and the affinity index (P < 0.05) were moderately higher around Ti/CS implants as compared to uncoated Ti. The highest values were measured in the G-Ti/CS and E-Ti/CS groups, the lowest values for the E-Ti and G-Ti controls. Quantitative synchrotron radiation micro-computed tomography (SRμCT) revealed the highest increase in total bone formation around G-Ti/CS as compared to C-Ti (P < 0.01). The effects with respect to direct bone apposition were less pronounced with SRμCT. Using scanning nanoindentation, both the indentation modulus and the hardness of the newly formed bone were highest in the E-Ti/CS, G-Ti/CS and G-Ti groups as compared to C-Ti (P < 0.05). Coatings with collagen and CS appear to improve both the quantity and quality of bone formed around Ti implants in ovarectomized rats. A simultaneous ethinyl estradiol- and genistein-rich diet seems to enhance these effects.     

Hierarchically structured titanium foams for tissue scaffold applications

We present a novel route for producing a new class of titanium foams for use in biomedical implant applications. These foams are hierarchically porous, with both the traditional large (>300 μm) highly interconnected pores and, uniquely, wall struts also containing micron scale (0.5–5 μm) interconnected porosities. The fabrication method consists of first producing a porous oxide precursor via a gel casting method, followed by electrochemical reduction to produce a metallic foam. This method offers the unique ability to tailor the porosity at several scales independently, unlike traditional space-holder techniques. Reducing the pressure during foam setting increased the macro-pore size. The intra-strut pore size (and percentage) can be controlled independently of macro-pore size by altering the ceramic loading and sintering temperature during precursor production. Typical properties for an 80% porous Ti foam were a modulus of ～1 GPa, a yield strength of 8 MPa and a permeability of 350 Darcies, all of which are in the range required for biomedical implant applications. We also demonstrate that the micron scale intra-strut porosities can be exploited to allow infiltration of bioactive materials using a novel bioactive silica–polymer composite, resulting in a metal–bioactive silica–polymer composite.