New Aspects in Osteoinduction

Bone grafting is a widely used procedure to treat bone defects. Various synthetic bone substitutes are gaining upon autologous cancellous bone, which is still the most effective graft material, despite the associated morbidity at the harvesting site. The lack of osteoinductive or osteogenic properties limits the success of those materials compared to autograft. One approach to enhance their biological behaviour is the addition of blood or bone marrow derived cellular material. Both platelet rich plasma (PRP) and fresh bone marrow (BM) have been reported to enhance bone formation clinically. The aim of this work was to evaluate in a pilot study the osteogenic potential of autologous substances like blood and bone marrow processed intra‐operatively in order to be impregnated on a synthetic ceramic bone substitute.     

A quantitative technique for comparing synthetic porous hydroxyapatite structures and cancellous bone

There are many different materials currently available for cancellous bone grafting. There is however, very little information relating the morphology of these materials to cancellous bone. Work was undertaken to develop a quantitative method for comparing synthetic hydroxyapatite bone structures with cancellous bone. The bases for comparison were mean plate thickness, mean plate distance, mineral area fraction, mineral volume fraction and plate orientation coupled with mechanical tests. The aim of this work was to develop a protocol for assessing whether these critical parameters which influence the success of bone implants were achieved in the synthetic materials. The methodology is successful in providing quantitative information about the mineral area fraction, the mean plate distance or pore size and the intercept frequency as a function of angle. Combining these three provides a quantitative measure of how much mineral there is and how it is distributed and oriented. The mechanical tests yield strengths and moduli values based on apparent density. The results of the mechanical tests can also be plotted as functions of the more discrete structural features such as those quantified in the image analysis to allow for even more equitable and systematic comparisons of different porous materials.     

Mechanical properties of open-cell foam synthetic thoracic vertebrae

This study presents comprehensive morphological and mechanical properties (static, dynamic) of open-cell rigid foams (Pacific Research Laboratories Inc. Vashon, WA) and a synthetic vertebral body derived from each of the foams. Synthetic vertebrae were comprised of a cylindrical open-cell foam core enclosed by a fiberglass resin cortex. The open-cell rigid foam was shown to have similar morphology and porosity as human vertebral cancellous bone, and exhibited a crush or fracture consolidation band typical of open-celled materials and cancellous bone. However, the foam material density was 40% lower than natural cancellous bone resulting in a lower compressive apparent strength and apparent modulus in comparison to human bone. During cyclic, mean compression fatigue tests, the synthetic vertebrae exhibited an initial apparent modulus, progressive modulus reduction, strain accumulation and S-N curve behaviour similar to human and animal vertebral cancellous bone. Synthetic open-cell foam vertebrae offer researchers an alternative to human vertebral bone for static and dynamic biomechanical experiments, including studies examining the effects of cement injection. Presented, in part, at the XXth Congress of the International Society of Biomechanics and 29th Annual Meeting of the American Society of Biomechanics, Cleveland, OH, July 31-August 5, 2005     

Fatigue characterization of a polymer foam to use as a cancellous bone analog material in the assessment of orthopaedic devices

Analog materials are used as a substitute to cancellous bone for in vitro biomechanical tests due to their uniformity, consistency in properties and availability. To date, only the static material properties of these materials have been assessed, although they are often used in fatigue tests. Cancellous bone exhibits complex material behavior when subjected to fatigue loads, including modulus degradation, accumulation of permanent strain and increasing hysteresis. Analog materials should exhibit similar fatigue behavior to cancellous bone if they are to be used in cyclic loading tests. In our study, a polymer foam (commercial name HEREX C70.55) has been studied for its static and fatigue behavior and compared with that of cancellous bone. In compression, the foam exhibited qualitatively similar mechanical behavior, but the degree of modulus degradation and accumulation of permanent strain was lower than expected for cancellous bone. In general, the tensile properties of the foam were greater than found in compression, the opposite to the mechanical behavior of cancellous bone. The methodology employed here could form the basis of selecting suitable analog materials for cancellous bone in the future.     

Optimizing a hydroxyapatite/tricalcium-phosphate ceramic as a bone graft extender for impaction grafting

The mechanical properties of morsellized bone allografts and synthetic hydroxapatite/tricalcium-phosphate (HA/TCP) ceramic extender materials for the use in impaction grafting revision hip surgery were investigated using two test methods: a basic compression test and an endurance test in an in-vitro model of an impaction grafted femur. Formalin fixed ovine bone graft was identified as mechanically similar to fresh human bone and thus suitable as an experimental material for in-vitro testing. For 1 : 1 volumetric mixes of bone allograft and synthetic extender, the granular ceramic's properties were varied in porosity, chemical composition, sintering temperature and particle size. Initial mechanical stability, a crucial prerequisite for clinical success in impaction grafting, was increased for all bone/extender mixes. A high porosity, tricalcium-phosphate rich ceramic of medium particle size and sintered at high temperatures was recognized as an optimized extender material for impaction grafting balancing the mechanical and biological demands. Using the extender without bone graft as a pure replacement is not recommended.     

High-strength mineralized collagen artificial bone

Mineralized collagen （MC） is a biomimetic material that mimics natural bone matrix in terms of both chemical composition and microstructure. The biomimetic MC possesses good biocompatibility and osteogenic activity, and is capable of guiding bone regeneration as being used for bone defect repair. However, mechanical strength of existing MC artificial bone is too low to provide effective support at human load-bearing sites, so it can only be used for the repair at non-load-bearing sites, such as bone defect filling, bone graft augmentation, and so on. In the present study, a high strength MC artificial bone material was developed by using collagen as the template for the biomimetic mineralization of the calcium phosphate, and then followed by a cold compression molding process with a certain pressure. The appearance and density of the dense MC were similar to those of natural cortical bone, and the phase composition was in conformity with that of animal＇s cortical bone demonstrated by XRD. Mechanical properties were tested and results showed that the compressive strength was comparable to human cortical bone, while the compressive modulus was as low as human cancellous bone. Such high strength was able to provide effective mechanical support for bone defect repair at human load-bearing sites, and the low compressive modulus can help avoid stress shielding in the application of bone regeneration. Both in vitro cell experiments and in v/vo implantation assay demonstrated good biocompatibility of the material, and in v/vo stability evaluation indicated that this high-strength MC artificial bone could provide long-term effective mechanical support at human load- bearing sites.     

Assessment of Bonelike® graft with a resorbable matrix using an animal model

Synthetic bone grafts have been developed to provide an alternative to autografts and allografts. Bonelike® is a patented synthetic osteoconductive bone graft that mimics the mineral composition of natural bone. In the present preliminary animal studies a user-friendly version of synthetic bone graft Bonelike® have been developed by using a resorbable matrix, Floseal®, as a vehicle and raloxifene hydrochloride as a therapeutic molecule, that is known to decrease osteoclast activity and therefore enhanced bone formation. From histological and scanning electron microscopy evaluations, the use of Bonelike® associated with Floseal® and raloxifene hydrochloride showed that new bone was rapidly apposed on implanted granules and also that the presence of the matrix and therapeutic molecule does not alter the proven highly osteoconductivity properties of Bonelike®. Therefore, this association may be one step-forward for the clinical applications of Bonelike® scaffolds since it is much more easy-to-handle when compared to granular materials.     

Synthesis and characterization of mechanically strong carboxymethyl cellulose–gelatin–hydroxyapatite nanocomposite for load-bearing orthopedic application

Novel three-dimensional hybrid polymer–hydroxyapatite nanocomposites have been developed as load-bearing synthetic bone graft through in situ mineralization process, using natural polymers carboxymethyl cellulose (CMC) and gelatin (Gel) as matrix. This process is simple and does not involve any chemical cross-linker. Detailed structural and physicochemical characterization of the samples disclosed that incorporation of gelatin with CMC assists the formation of CMC-Gel polymeric network of new conformational structure through non-covalent interactions (H-bond). The formation of hydroxyapatite (HA) in this polymeric network was occurred in such a fashion that the HA serves as bridging molecule which strengthen the polymeric network more and formed a mechanically strong three-dimensional CMC-Gel-HA nanocomposite. The synthesized CMC-Gel-HA nanocomposites have compressive strength and modulus in the range of 40–86 MPa and 0.4–1.2 GPa, respectively, analogous to human cancellous as well as cortical bone. In vitro cell interaction of the synthesized nanocomposites with osteoblast-like MG-63 cells has been evaluated. Results showed that synthesized CMC-Gel-HA nanocomposite promote cells for high alkaline phosphatase activity and extracellular mineralization. Extracellular mineralization ability of nanocomposite was investigated by alizarin red staining and von Kossa staining. Biodegradable nature and bone apatite formation ability of CMC-Gel-HA nanocomposite under simulated physiological environment were investigated by different characterization processes. Results indicated that the synthesized CMC-Gel-HA nanocomposite has great potential to be used as regenerative bone graft in major load-bearing region.     

A new sheep model with automatized analysis of biomaterial-induced bone tissue regeneration

Presently, several bone graft substitutes are being developed or already available for clinical use. However, the limited number of clinical and in vivo trials for direct comparison between these products may complicate this choice. One of the main reasons for this scarcity it is the use of models that do not readily allow the direct comparison of multiple bone graft substitutes, due especially to the small number of implantation sites. Although sheep cancellous bone models are now well established for these purposes, the limited availability of cancellous bone makes it difficult to find multiple comparable sites within a same animal. These limitations can be overcome by the monocortical model here proposed as it consists in 5–6 holes (5 mm Ø), in the femoral diaphysis, with similar bone structure, overlying soft tissue and loading pattern for all defects. Associated to this model, it is also described a fast histomorphometric analysis method using a computer image segmentation test (Threshold method) to assess bone regeneration parameters. The information compiled through the experimental use of 45 sheep in several studies allowed determining that this ovine model has the potential to demonstrate differences in bone-forming performance between various scaffolds. Additionally, the described histomorphometric method is fast, accurate and reproducible.     

The processing and characterization of animal-derived bone to yield materials with biomedical applications. Part III: material and mechanical properties of fresh and processed bovine cancellous bone

Conversion of bovine cancellous bone to a useful biomedical xenograft material involves several processing steps which include boiling, defatting and deproteination (i.e. bleaching). This study has shown how these processes can influence cancellous bone modulus and strength. It was found that prolonged boiling in water for six hours followed by NaOCI bleaching had a deleterious effect on the overall strength of the bovine bone. In contrast, bone samples subjected to only moderate boiling (1.5 hours) exhibited a 22% stiffness increase due mainly to the effects of drying. The same stiffened samples, when subjected to the bleaching procedure, retained some strength with only a small reduction in moduli values. It can be concluded that careful control of defatting and bleaching procedures on bovine bone is able to give a strong, albeit, brittle material with preservation of the original bone architecture. The bone xenograft materials are worthy of further investigation in in vivo clinical trials to assess their performance in contact with biological fluids. © 2000 Kluwer Academic Publishers     

Properties of open-cell porous metals and alloys for orthopaedic applications

One shortcoming of metals and alloys used to fabricate various components of orthopaedic systems, such as the femoral stem of a total hip joint replacement and the tibial plate of a total knee joint replacement, is well-recognized. This is that the material modulus of elasticity (E′) is substantially larger than that of the contiguous cancellous bone, a consequence of which is stress shielding which, in turn, has been postulated to be implicated in a cascade of events that culminates in the principal life-limiting phenomenon of these systems, namely, aseptic loosening. Thus, over the years, a host of research programs have focused on the synthesis of metallic biomaterials whose E′ can be tailored to match that of cancellous bone. The present work is a review of the extant large volume of literature on these materials, which are called open-cell porous metals/alloys (or, sometimes, metal foams or cellular materials). As such, its range is wide, covering myriad aspects such as production methods, characterization studies, in vitro evaluations, and in vivo performance. The review also includes discussion of seven areas for future research, such as parametric studies of the influence of an assortment of process variables (such as the space holder material and the laser power in the space holder method and the laser-engineered net-shaping process, respectively) on various properties (notably, permeability, fatigue strength, and corrosion resistance) of a given porous metal/alloy, innovative methods of determining fatigue strength, and modeling of corrosion behavior.     

Bone remodelling in the pores and around load bearing transchondral isoelastic porous-coated glassy carbon implants: Experimental study in rabbits

Cylinders of porous-coated glassy carbon were implanted into drill holes made through the articular surface of the medial condyle of both tibiae of ten rabbits for six and 12 weeks. Bone ingrowth and remodelling was examined by radiographic, histologic, oxytetracycline-fluorescence and microradiographic methods. Bone ingrowth into pores and load bearing implants was seen by all examination methods. Bone ingrowth occurred earlier when the pores were facing cancellous bone than cortical bone. Appositional bone formation occurred on the trabeculae a few millimetres from the interface during the early phase of remodelling at six weeks. At 12 weeks resorptive remodelling had occurred both in the surroundings and in those pores that face cancellous bone, whereas the amount of bone still increased in the pores facing cortical bone. In its porous-coated form glassy carbon functions well as a frame for ingrowing bone and it shows good osteoconductivity. Its mechanical properties are suitable for functioning as a structural bone substitute in places where the loads are mainly compressive. The difference between findings at six and 12 weeks indicated physiologic stress distribution and the adverse effects of stiff materials on bone remodelling were avoided by using this isoelastic material.     

Hydrogel/bioactive glass composites for bone regeneration applications: Synthesis and characterisation

Due to the deficiencies of current commercially available biological bone grafts, alternative bone graft substitutes have come to the forefront of tissue engineering in recent times. The main challenge for scientists in manufacturing bone graft substitutes is to obtain a scaffold that has sufficient mechanical strength and bioactive properties to promote formation of new tissue. The ability to synthesise hydrogel based composite scaffolds using photopolymerisation has been demonstrated in this study. The prepared hydrogel based composites were characterised using techniques including Fourier Transform Infrared Spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), Energy-dispersive X-ray spectrometry (EDX), rheological studies and compression testing. In addition, gel fraction, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), porosity and swelling studies of the composites were carried out. It was found that these novel hydrogel bioglass composite formulations did not display the inherent brittleness that is typically associated with bioactive glass based bone graft materials and exhibited enhanced biomechanical properties compared to the polyethylene glycol hydrogel scaffolds along. Together, the combination of enhanced mechanical properties and the deposition of apatite on the surface of these hydrogel based composites make them an ideal candidate as bone graft substitutes in cancellous bone defects or low load bearing applications.     

In vitro evaluation of bioactive strontium-based ceramic with rabbit adipose-derived stem cells for bone tissue regeneration

The development of bone replacement materials is an important objective in the field of orthopaedic surgery. Due to the drawbacks of treating bone defects with autografts, synthetic bone graft materials have become optional. So in this work, a bone tissue engineering approach with radiopaque bioactive strontium incorporated calcium phosphate was proposed for the preliminary cytocompatibility studies for bone substitutes. Accumulating evidence indicates that strontium containing biomaterials promote enhanced bone repair and radiopacity for easy imaging. Hence, strontium calcium phosphate (SrCaPO₄) and hydroxyapatite scaffolds have been investigated for its ability to support and sustain the growth of rabbit adipose-derived mesenchymal stem cells (RADMSCs) in vitro. They were characterized via Micro-CT for pore size distribution. Cells used were isolated from New Zealand White rabbit adipose tissue, characterized by FACS and via differentiation into the osteogenic lineage by alkaline phosphatase, Masson’s trichome, Alizarin Red and von Kossa staining on day 28. Material-cell interaction was observed by SEM imaging of cell morphology on contact with material. Live–Dead analysis was done by confocal laser scanning microscopy and cell cluster analysis via μCT. The in vitro biodegradation, elution and nucleation of apatite formation of the material was evaluated using simulated body fluid and phosphate buffered saline in static regime up to 28 days at 37 °C. These results demonstrated that SrCaPO₄ is a good candidate for bone tissue engineering applications and with osteogenically-induced RADMSCs, they may serve as potential implants for the repair of critical-sized bone defects.     

Bioinspired Materials with Self-Adaptable Mechanical Properties

Natural structural materials, such as bone, can autonomously modulate their mechanical properties in response to external loading to prevent failure. These material systems smartly control the addition/removal of material in locations of high/low mechanical stress by utilizing local resources guided by biological signals. On the contrary, synthetic structural materials have unchanging mechanical properties limiting their mechanical performance and service life. Inspired by the mineralization process of bone, a material system that adapts its mechanical properties in response to external mechanical loading is reported. It is found that charges from piezoelectric scaffolds can induce mineralization from surrounding media. It is shown that the material system can adapt to external mechanical loading by inducing mineral deposition in proportion to the magnitude of the stress and the resulting piezoelectric charges. Moreover, the mineralization mechanism allows a simple one-step route for fabricating functionally graded materials by controlling the stress distribution along the scaffold. The findings can pave the way for a new class of self-regenerating materials that reinforce regions of high stress or induce deposition of minerals on the damaged areas from the increase in mechanical stress to prevent/mitigate failure. It is envisioned that the findings can contribute to addressing the current challenges of synthetic materials for load-bearing applications from self-adaptive capabilities.     

Constraining the lateral dimensions of uniaxially loaded materials increases the calculated strength and stiffness: application to muscle and bone

If a solid body is deformed along one direction, by a uniaxial applied stress for instance, then strains will also be induced in perpendicular directions. The negative ratio of the induced strain to the applied strain is known as the Poisson ratio. Analysis of the elasticity tensor relating stress and strain within a solid shows that if the induced strain is restricted, then a greater stress is required to produce the same strain; it appears stiffer. Many biological materials with a mechanical function are subject to forces which are primarily uniaxial. This mechanism appears to be used to maximize the uniaxial load-bearing properties of some of these materials. Muscles are commonly surrounded by strong sheets of connective tissue which will constrain the lateral expansion of the muscle as it contracts. This increases the stress in the muscle for a given strain, and hence the load it can support. Similarly, cancellous bone is normally surrounded by a shell of much stronger compact bone and this effectively increases the stiffness of the cancellous bone without the penalty of increasing the mass, as would be the case if the same stiffening was produced by increasing the degree of calcification. It also has important implications for the failure of bone, which is largely a function of strain rather than stress.     

Minimum solid area models applied to the prediction of Young's modulus for cancellous bone

In developing models for the mechanical behavior of cancellous bone, accurate prediction of Young's modulus as a function of the pore fraction and morphology is a requirement. Previous workers have suggested models which provide good statistical fits, but most of these models are highly idealized, with no treatment of the actual morphology of the porosity. In the field of engineering ceramics, simple minimum solid area models have been developed over the past four decades to describe the mechanical properties of porous structural ceramics. This paper applies these models to data for cancellous bone, and it is shown that one, developed specifically for high porosity materials, gives realistic predictions of tissue modulus and a good statistical fit to well-established data. This model should prove to be useful in biomechanical analyses involving cancellous bone tissue.     

Review Micromechanical testing of bone trabeculae - potentials and limitations

The mechanical properties of bone are studied mostly for reasons related to skeletal pathology. However, bone is also very interesting from a material science perspective because it is a natural hierarchical composite material. The mechanical properties of bone depend on both the structural arrangement and the properties of the constituting materials, namely the organic polymer collagen and the inorganic salt apatite. While the mechanical properties of bone samples at the macroscopic scale are measured routinely, mechanical tests on micrometer-sized specimens are still at development stage. In this paper, protocols for measuring the elasticity of cancellous bone trabeculae are reviewed. The published values for the elastic modulus of trabeculae vary between 1 GPa and 15 GPa. Reasons for this broad range of values may be located in the intrinsic difficulties of preparing, handling, and testing inhomogeneous, anisotropic and asymmetric micro-samples. We discuss the major error sources in existing testing procedures and suggest potential strategies to enhance their performance.     

Determination of mechanical properties of impacted human morsellized cancellous allografts for revision joint arthroplasty

This paper deals with the characterization of mechanical properties of impacted morsellized cancellous allograft (IMCA) produced by dynamic compaction of allograft femoral heads ground by commercially available bone mills, i.e. rotating rasp and reciprocating type bone mills. Various ranges and profiles of particle size in the graft aggregates were obtained using these bone mills, and the effect of number of compaction as well as the distribution of particle sizes on the mechanical properties of IMCA under quasistatic compression and shear loading conditions was discussed. The morsellized cancellous allograft prepared by the reciprocating type bone mill showed a broad distribution of particle sizes, and gave IMCA superior mechanical properties to the graft with a more uniform size distribution, or prepared by the rotating rasp type bone mills. The increase of number of compaction also improved the mechanical properties of IMCA in compression.     

Klinische Anwendungsmöglichkeiten von porösen Biomaterialien im Knochengewebe

Clinical applications of porous biomaterials in bone During therapy of bone defects and lesions it often exists the necessity to treat larger defects. While doing so porous biomaterials are frequently used as an alternative to autologeous bone in the field of implantology, traumatology and orthopaedic surgery with good clinical results. In contrast to autologeous cancellous bone grafts they are of an almost unlimited availability and are not afflicted with the problem of donor morbidity. In order to accelerate the quantity and quality of the newly formed bone and thus to enable the reconstruction of a defect, different biomaterials are used as bone substitutes. These are materials which are able to interact with biological systems, and mostly synthetic porous tricalciumphophates and hydroxyapatite ceramics as well as processed allografts are used. As an additional possibility of clinical application surface coating of implants with porous biomaterials and bone‐forming growth factors comes into consideration. In the presented work an overview of the clinical application possibilities of porous biomaterials in bone as well as an outlook on future developments is supposed to be given.