The effect of scaffold-cell entrapment capacity and physico-chemical properties on cartilage regeneration.

An important tenet in designing scaffolds for regenerative medicine consists in mimicking the dynamic mechanical properties of the tissues to be replaced to facilitate patient rehabilitation and restore daily activities. In addition, it is important to determine the contribution of the forming tissue to the mechanical properties of the scaffold during culture to optimize the pore network architecture. Depending on the biomaterial and scaffold fabrication technology, matching the scaffolds mechanical properties to articular cartilage can compromise the porosity, which hampers tissue formation. Here, we show that scaffolds with controlled and interconnected pore volume and matching articular cartilage dynamic mechanical properties, are indeed effective to support tissue regeneration by co-cultured primary and expanded chondrocyte (1:4). Cells were cultured on scaffolds in vitro for 4 weeks. A higher amount of cartilage specific matrix (ECM) was formed on mechanically matching (M) scaffolds after 28 days. A less protein adhesive composition supported chondrocytes rounded morphology, which contributed to cartilaginous differentiation. Interestingly, the dynamic stiffness of matching constructs remained approximately at the same value after culture, suggesting a comparable kinetics of tissue formation and scaffold degradation. Cartilage regeneration in matching scaffolds was confirmed subcutaneously in vivo. These results imply that mechanically matching scaffolds with appropriate physico-chemical properties support chondrocyte differentiation.

Bulk properties and bioactivity assessment of porous polymethylmethacrylate cement loaded with calcium phosphates under simulated physiological conditions.

Polymethylmethacrylate (PMMA) cements are widely used in spinal surgery. Nevertheless, these types of cements present some documented drawbacks. Therefore, efforts have been made to improve the properties and biological performance of solid PMMA. A porous structure would seem to be advantageous for anchoring purposes. This work studied the bulk physicochemical, mechanical and interconnectivity properties of porous PMMA cements loaded with various amounts of calcium phosphate (CaP). As a measure of bioactivity, changes of PMMA cements under simulated physiological conditions were studied in a calcium phosphate solution for 0, 3, 7, 14, 21 and 28 days. Scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), micro-computed tomography (µ-CT) and mechanical compression tests were performed to characterize the morphology, crystallographic and chemical composition, interconnectivity and mechanical properties, respectively. SEM allowed observing the result of loading CaP into the porous PMMA, which was corroborated by XRD, FTIR and µ-CT. No interference of the CaP with the PMMA was detected. µ-CT described similar interconnectivity and pore distribution for all CaP percentages. Mechanical properties were not significantly altered by the CaP percentages or the immersion time. Hence, porous PMMA was effectively loaded with CaP, which provided the material with properties for potential osteoconductivity.

The effect of scaffold architecture on properties of direct 3D fiber deposition of porous Ti6Al4V for orthopedic implants.

3D porous Ti6Al4V scaffolds were directly fabricated by a rapid prototyping technology, 3D fiber deposition (3DF). In this study, scaffolds with different structures were fabricated by changing fiber spacing and fiber orientation. The influence of different architectures on mechanical properties and permeability of the scaffold were investigated. Mechanical analysis revealed that compressive strength and E-modulus increase with decreasing the porosity. Permeability measurements showed that not only the total porosity but also the porous structure can influence the permeability. 3DF was found to provide good control and reproducibility of the desired degree of porosity and the 3D structure. Results of this study demonstrate that the 3DF of Ti6Al4V give us flexibility and versatility to fabricate and improve scaffolds to better mimic the architecture and properties of natural bone and meet the requirements of bone graft substitutes and orthopedic and dental implants.

Integrating novel technologies to fabricate smart scaffolds.

Tissue engineering aims at restoring or regenerating a damaged tissue by combining cells, derived from a patient biopsy, with a 3D porous matrix functioning as a scaffold. After isolation and eventual in vitro expansion, cells are seeded on the 3D scaffolds and implanted directly or at a later stage in the patient's body. 3D scaffolds need to satisfy a number of requirements: (i) biocompatibility, (ii) biodegradability and/or bioresorbability, (iii) suitable mechanical properties, (iv) adequate physicochemical properties to direct cell-material interactions matching the tissue to be replaced and (v) ease in regaining the original shape of the damaged tissue and the integration with the surrounding environment. Still, it appears to be a challenge to satisfy all the aforementioned requisites with the biomaterials and the scaffold fabrication technologies nowadays available. 3D scaffolds can be fabricated with various techniques, among which rapid prototyping and electrospinning seem to be the most promising. Rapid prototyping technologies allow manufacturing scaffolds with a controlled, completely accessible pore network--determinant for nutrient supply and diffusion--in a CAD/CAM fashion. Electrospinning (ESP) allows mimicking the extracellular matrix (ECM) environment of the cells and can provide fibrous scaffolds with instructive surface properties to direct cell faith into the proper lineage. Yet, these fabrication methods have some disadvantages if considered alone. This review aims at summarizing conventional and novel scaffold fabrication techniques and the biomaterials used for tissue engineering and drug-delivery applications. A new trend seems to emerge in the field of scaffold design where different scaffolds fabrication technologies and different biomaterials are combined to provide cells with mechanical, physicochemical and biological cues at the macro-, micro- and nano-scale. If merged together, these integrated technologies may lead to the generation of a new set of 3D scaffolds that satisfies all of the scaffolds' requirements for tissue-engineering applications and may contribute to their success in a long-term scenario.

Design of biphasic polymeric 3-dimensional fiber deposited scaffolds for cartilage tissue engineering applications.

This report describes a novel system to create rapid prototyped 3-dimensional (3D) fibrous scaffolds with a shell-core fiber architecture in which the core polymer supplies the mechanical properties and the shell polymer acts as a coating providing the desired physicochemical surface properties. Poly[(ethylene oxide) terephthalate-co-poly(butylene) terephthalate] (PEOT/PBT) 3D fiber deposited (3DF) scaffolds were fabricated and examined for articular cartilage tissue regeneration. The shell polymer contained a higher molecular weight of the initial poly(ethylene glycol) (PEG) segments used in the copolymerization and a higher weight percentage of the PEOT domains compared with the core polymer. The 3DF scaffolds entirely produced with the shell or with the core polymers were also considered. After 3 weeks of culture, scaffolds were homogeneously filled with cartilage tissue, as assessed by scanning electron microscopy. Although comparable amounts of entrapped chondrocytes and of extracellular matrix formation were found for all analyzed scaffolds, chondrocytes maintained their rounded shape and aggregated during the culture period on shell-core 3DF scaffolds, suggesting a proper cell differentiation into articular cartilage. This finding was also observed in the 3DF scaffolds fabricated with the shell composition only. In contrast, cells spread and attached on scaffolds made simply with the core polymer, implying a lower degree of differentiation into articular cartilaginous tissue. Furthermore, the shell-core scaffolds displayed an improved dynamic stiffness as a result of a "prestress" action of the shell polymer on the core one. In addition, the dynamic stiffness of the constructs increased compared with the stiffness of the bare scaffolds before culture. These findings suggest that shell-core 3DF PEOT/PBT scaffolds with desired mechanical and surface properties are a promising solution for improved cartilage tissue engineering.

[Dissertations 25 years after date 14. The development of porous polymethylmethacrylate cement as potential implant material]. / Proefschriften 25 jaar na dato 14 De ontwikkeling van een poreus polymethylmethacrylaat cement als botvervangend materiaal.

The development of polymethylmethacrylate bone cement with an open spongelike structure resulted as a spin-off from efforts to dampen the high exothermal peak during hardening of the cement. A cement formulation in which an aqueous gel was mixed through the organic dough showed the desired significantly reduced temperature peak and improved biocompatibility but also ingrowth of bone into the pores that had been formed by the resorbing aqueous gel in the polymerized matrix. The expectation that such a cement would provide a better fixation of total hip prostheses did not come true because of the diminished mechanical strength due to the same porosity. Better applicability of the cement was found for bony areas that were not subjected to heavy stresses such as bone defect filling and augmentation of cranio-facial and sternal deformities. In spite of successful clinical trials and positive findings after long term evaluations a commercial development of the cement was not undertaken because of a market that was estimated too small to be profitable.

Finite Element Analysis of Meniscal Anatomical 3D Scaffolds: Implications for Tissue Engineering.

Solid Free-Form Fabrication (SFF) technologies allow the fabrication of anatomical 3D scaffolds from computer tomography (CT) or magnetic resonance imaging (MRI) patients' dataset. These structures can be designed and fabricated with a variable, interconnected and accessible porous network, resulting in modulable mechanical properties, permeability, and architecture that can be tailored to mimic a specific tissue to replace or regenerate. In this study, we evaluated whether anatomical meniscal 3D scaffolds with matching mechanical properties and architecture are beneficial for meniscus replacement as compared to meniscectomy. After acquiring CT and MRI of porcine menisci, 3D fiber-deposited (3DF) scaffolds were fabricated with different architectures by varying the deposition pattern of the fibers comprising the final structure. The mechanical behaviour of 3DF scaffolds with different architectures and of porcine menisci was measured by static and dynamic mechanical analysis and the effect of these tissue engineering templates on articular cartilage was assessed by finite element analysis (FEA) and compared to healthy conditions or to meniscectomy. Results show that 3DF anatomical menisci scaffolds can be fabricated with pore different architectures and with mechanical properties matching those of natural menisci. FEA predicted a beneficial effect of meniscus replacement with 3D scaffolds in different mechanical loading conditions as compared to meniscectomy. No influence of the internal scaffold architecture was found on articular cartilage damage. Although FEA predictions should be further confirmed by in vitro and in vivo experiments, this study highlights meniscus replacement by SFF anatomical scaffolds as a potential alternative to meniscectomy.

Dynamic mechanical properties of 3D fiber-deposited PEOT/PBT scaffolds: an experimental and numerical analysis.

Mechanical properties of three-dimensional (3D) scaffolds can be appropriately modulated through novel fabrication techniques like 3D fiber deposition (3DF), by varying scaffold's pore size and shape. Dynamic stiffness, in particular, can be considered as an important property to optimize the scaffold structure for its ultimate in vivo application to regenerate a natural tissue. Experimental data from dynamic mechanical analysis (DMA) reveal a dependence of the dynamic stiffness of the scaffold on the intrinsic mechanical and physicochemical properties of the material used, and on the overall porosity and architecture of the construct. The aim of this study was to assess the relationship between the aforementioned parameters, through a mathematical model, which was derived from the experimental mechanical data. As an example of how mechanical properties can be tailored to match the natural tissue to be replaced, articular bovine cartilage and porcine knee meniscus cartilage dynamic stiffness were measured and related to the modeled 3DF scaffolds dynamic stiffness. The dynamic stiffness of 3DF scaffolds from poly(ethylene oxide terephthalate)-poly(butylene terephthalate) (PEOT/PBT) copolymers was measured with DMA. With increasing porosity, the dynamic stiffness was found to decrease in an exponential manner. The influence of the scaffold architecture (or pore shape) and of the molecular network properties of the copolymers was expressed as a scaffold characteristic coefficient alpha, which modulates the porosity effect. This model was validated through an FEA numerical simulation performed on the structures that were experimentally tested. The relative deviation between the experimental and the finite element model was less than 15% for all of the constructs with a dynamic stiffness higher than 1 MPa. Therefore, we conclude that the mathematical model introduced can be used to predict the dynamic stiffness of a porous PEOT/PBT scaffold, and to choose the biomechanically optimal structure for tissue engineering applications.

3D fiber-deposited scaffolds for tissue engineering: influence of pores geometry and architecture on dynamic mechanical properties.

One of the main issues in tissue engineering is the fabrication of scaffolds that closely mimic the biomechanical properties of the tissues to be regenerated. Conventional fabrication techniques are not sufficiently suitable to control scaffold structure to modulate mechanical properties. Within novel scaffold fabrication processes 3D fiber deposition (3DF) showed great potential for tissue engineering applications because of the precision in making reproducible 3D scaffolds, characterized by 100% interconnected pores with different shapes and sizes. Evidently, these features also affect mechanical properties. Therefore, in this study we considered the influence of different structures on dynamic mechanical properties of 3DF scaffolds. Pores were varied in size and shape, by changing fibre diameter, spacing and orientation, and layer thickness. With increasing porosity, dynamic mechanical analysis (DMA) revealed a decrease in elastic properties such as dynamic stiffness and equilibrium modulus, and an increase of the viscous parameters like damping factor and creep unrecovered strain. Furthermore, the Poisson's ratio was measured, and the shear modulus computed from it. Scaffolds showed an adaptable degree of compressibility between sponges and incompressible materials. As comparison, bovine cartilage was tested and its properties fell in the fabricated scaffolds range. This investigation showed that viscoelastic properties of 3DF scaffolds could be modulated to accomplish mechanical requirements for tailored tissue engineered applications.

Porous Ti6Al4V scaffolds directly fabricated by 3D fibre deposition technique: effect of nozzle diameter.

3D porous Ti6Al4V scaffolds were successfully directly fabricated by a rapid prototyping technology: 3D fibre deposition. In this study, the rheological properties of Ti6Al4V slurry was studied and the flow rate was analyzed at various pressures and nozzle diameters. Scaffolds with different fibre diameter and porosity were fabricated. ESEM observation and mechanical tests were performed on the obtained porous Ti6Al4V scaffolds with regard to the porous structure and mechanical properties. The results show that these scaffolds have 3D interconnected porous structure and a compressive strength which depends on porosity at constant fibre diameters and on the fibre diameter at constant porosity. These Ti6Al4V scaffolds are expected to be constructs for biomedical applications.

Three-dimensional fiber-deposited PEOT/PBT copolymer scaffolds for tissue engineering: influence of porosity, molecular network mesh size, and swelling in aqueous media on dynamic mechanical properties.

Among novel scaffold fabrication techniques, 3D fiber deposition (3DF) has recently emerged as a means to fabricate well-defined and custom-made scaffolds for tissue regeneration, with 100% interconnected pores. The mechanical behavior of these constructs is dependent not only on different three-dimensional architectural and geometric features, but also on the intrinsic chemical properties of the material used. These affect the mechanics of the solid material and eventually of 3D porous constructs derived from them. For instance, poly(ethylene oxide terephthalate)-poly(butylene terephthalate) (PEOT/PBT) block copolymers are known to have mechanical properties, depending on the PEOT/PBT weight ratio in block form and on the molecular weight of the initial poly(ethylene glycol) (PEG) blocks. These differences are enhanced even more by their different swelling properties in aqueous media. Therefore, this article examines the influence of copolymer compositions in terms of their swelling on dynamic mechanical properties of solid material and porous 3DF scaffolds. The molecular weight of the starting PEG blocks used in the copolymer synthesis varied from 300 to 1000 g/mol. The PEOT/PBT weight ratio in the blocks used varied from 55/45 to 80/20. This corresponded to an increase of the swelling ratio Q from 1.06 to 2.46, and of the mesh size xi from approximately 9 Angstrom to approximately 47 Angstrom. With increased swelling, dynamic mechanical analysis (DMA) revealed a decrease in elastic response and an increase of viscoelasticity. Thus, by coupling structural and chemical characteristics, the viscoelastic properties of PEOT/PBT 3DF scaffolds may be fine tuned to achieve mechanical requirements for a variety of engineered tissues. Ultimately, the combination of 3DF and DMA may be useful to validate the hypothesis that mimicking the biomechanical behavior of a specific tissue for its optimal replacement is an important issue for at least some tissue-engineering applications.

Use of bone-bonding hydrogel copolymers in bone: an in vitro and in vivo study of expanding PEO-PBT copolymers in goat femora.

Polyactive(R) [polyethylene oxide-polybuthylene terephtalate (PEO-PBT)] refers to a group of copolymers with bone-bonding properties. In reference to these properties, PEO-PBT copolymers are currently being investigated for their possible use in orthopedic surgery and dentistry. PEO-PBT copolymers exhibit hydrogel behavior. When swelling in fluid is prohibited by mechanical confinement, the copolymers exert a swelling pressure on surrounding structures. In the first part of this study, these swelling pressures were measured in vitro. Polymers with different ratios of PEO-PBT exerted a swelling pressure of more than 2 MPa when tested in fluid between the cross-heads of a Hounsfield test-bench. In the second part of the study, the biocompatibility of PEO-PBT 55-45 and the effect of continuous intramedullary pressure of these copolymers on bone was investigated. Large cylinders of dry PEO-PBT 55-45 were implanted with a tight fit in the distal part of goat femora. Preswollen cylinders of PEO-PBT implanted in the opposite femur served as a control. Although it was assumed that the pressure of dry PEO-PBT on the bone would reach more than 2 MPa with press-fit insertion, no immediate hazardous effects of the expanding polymer were noticed within the first days postoperatively. The goats were sacrificed after 3, 9, and 25 weeks. Histological examination showed good implant-bone contact at different follow-up times in the distal femora with the dry implanted implants. The femora in which the preswollen cylinders had been implanted showed a thin layer of soft tissue between the PEO-PBT implant and bone. The swelling pressure exerted by dry press-fit implanted PEO-PBT implants is an important factor in creating a strong interface bond between PEO-PBT and bone.

Incorporation of bovine serum albumin in calcium phosphate coating on titanium.

Calcium phosphate (Ca-P) and bovine serum albumin (BSA) were coprecipitated as a coating on commercially pure titanium (cpTi) with a high protein loading (15 wt %) by employing a recently developed wet-chemistry technique. It was observed that the incorporation of BSA significantly modified the morphology, composition, and crystallinity of the Ca-P coating. The Ca-P coating without BSA is a mixture of hydroxyapatite (HA) and octacalcium phosphate (OCP) with sharp-edged thin OCP crystal plates on the top layer, whereas only an HA phase was detected in the Ca-P/BSA coating. The crystal plates in the latter had a more rounded appearance. The Ca-P/BSA coatings were immersed respectively in neutral (pH 7.4) and acidic (starting pH 4.0) phosphate-buffered saline (PBS) at 37 degrees C over a 14-day period. No protein release was detected in the neutral PBS during the immersion; however, a continuous release of BSA was measured in the acidic PBS, subsequently leading to the formation of a very dense and well-adherent composite coating of BSA and Ca-P on cpTi. The present study provides the possibility to achieve a long-term effective release of biologically active proteins from a Ca-P-coated metallic implant.

Surface modification of nano-apatite by grafting organic polymer.

Since surface properties of hydroxyapatite (HA) play an important role in its performance, surface modification of HA has gained much attention from researchers. Silane coupling agents have been the focus of the research. In this study, an effective surface modification method was developed using hexamethylene diisocyanate as a coupling agent. Polyethylene glycol (Mw = 1500) was successfully coupled to the surface of nano-size apatite particles (nano-apatite). Various methods were used to characterize the surface-modified nano-apatite. Infra-red spectra confirmed the existence of a layer of polymer with both urethane and ether linkage on the surface of nano-apatite. The amount of grafted polymer as determined by total organic carbon analysis (TOC) and thermal gravimetric analysis (TGA) was about 20% in weight. Solid 1H MAS NMR spectra indicated that the amount of hydroxyl groups of nano-apatite was decreased by 7.7% after surface grafting reaction. It is concluded that the surface hydroxyl groups of nano-apatite have the reactivity towards isocyanate groups.

Preparation of bioactive Ti6Al4V surfaces by a simple method.

Boiling diluted alkali incubation was found to be an effective way to prepare bioactive Ti6Al4V surfaces, whether polished or not, as indicated in vitro after immersion in two different supersaturated calcification solutions (SCSs). The induction of calcium phosphate (Ca-P) precipitation from the SCSs is most probably made possible by the formation of a new TiO2 surface layer and a large number of submicron-scaled etched pits therein. The morphologies and composition of the Ca-P deposited from different SCSs are entirely different from each other. The processes on Ti6Al4V surfaces during treatment and immersion were investigated in detail by means of scanning electron microscopy combined with energy dispersive X-ray analysis, X-ray photoelectron spectroscopy and X-ray diffraction.

Preparation of calcium phosphate coatings on titanium implant materials by simple chemistry.

A two-step chemical treatment has been developed in our group to prepare commercially pure titanium (cpTi) surfaces that will allow calcium phosphate (Ca-P) precipitation during immersion in a supersaturated calcification solution (SCS) with ion concentrations of [Ca2+] = 3.10 mM and [HPO4(2-)] = 1.86 mM. It was observed that a precalcification (Pre-Ca) procedure prior to immersion could significantly accelerate the Ca-P deposition process. In this work, the bioactivity of chemically treated cpTi and Ti6Al4V was further verified by applying commercially available Hanks' balanced salt solution (HBSS), an SCS with very low ion concentrations of [Ca2+] = 1.26 mM and [HPO4(2-)] = 0.779 mM, as the immersion solution. It was found that a uniform and very dense apatite coating magnesium impurities was formed if the Pre-Ca procedure was performed before immersion, as compared with the loose Ca-P layer obtained from the abovementioned high concentration of SCS. The formation of a microporous titanium dioxide thin surface layer on cpTi or Ti6Al4V by the two-step chemical treatment could be the main reason for the induction of apatite nucleation and growth from HBSS. Variations of pH values, Ca and P concentrations, and immersion time in HBSS were investigated to reveal the detailed process of Ca-P deposition. The described treatments provide a simple chemical method to prepare Ca-P coatings on both cpTi and Ti6Al4V.

Collagen/apatite coating on 3-dimensional carbon/carbon composite.

A three-dimensional carbon/carbon composite (3D C/C) was studied as potential bone-repairing material; its major mechanical properties were found to be closer to those of human bone than other common bone-repairing materials available. In vitro calcification tests revealed that as-received 3D C/C is almost bioinert in simulated body fluid (SBF) over an immersion period of 4 weeks. To improve the bioactivity of 3D C/C, surface modification was accomplished through two practical routes: (1) grafting with polyethylene glycol (PEG) and (2) phosphorylation and precalcification. After grafting with alpha, omega di(aminopropyl) polyethylene glycol 800 (NH2-PEG-NH2), a continuous layer of calcium phosphate was formed on the surface of 3D C/C in SBF after 4 weeks. Phosphorylated 3D C/C samples have the ability to induce apatite precipitation after precalcification in a saturated Ca(OH)2 solution for 1 week. To speed up the coating process, a calcification solution with collagen was developed in which a collagen/apatite coating layer can be formed on 3D C/C in 9 h in ambient conditions.

A study on the grafting reaction of isocyanates with hydroxyapatite particles.

The surface grafting reactions of a series of isocyanates with hydroxyapatite particles at different temperatures were studied by Infrared spectrophotometry (IR) and thermal gravimetric analysis (TGA). The study results show that both hexamethylene diisocyanate (HMDI) and isocyanatoethyl methacrylate (ICEM) react readily with HA while ethyl isocyanate acetate (EIA) and butyl isocyanate (BIC) have lower reactivity towards HA particles. It also has been found that the reaction of ICEM with HA follows a second-order reaction mechanism, despite the heterogeneous nature of the reaction, while the reaction of HMDI with HA does not due to the complexity of the reaction. Based on this study, it is concluded that ICEM and HMDI are suitable agents for the coupling of polymers due to their reactivity towards HA.

Composite biomaterials with chemical bonding between hydroxyapatite filler particles and PEG/PBT copolymer matrix.

In an effort to make composites from hydroxyapatite and a PEG/PBT copolymer (Polyactive 70/30), chemical linkages were introduced between the filler particles and polymer matrix using hexamethylene diisocyanate as a coupling agent. Infrared spectra (IR) and thermal gravimetric analysis (TGA) confirmed the presence of Polyactive 70/30 on the surface of HA filler particles. The amount of chemically bound polymer was 4.7 wt.%, as determined by TGA. The mechanical properties of the composites, that is, tensile strength and Young's modulus, were improved significantly by the introduction of a chemical linkage between the filler particles and polymer matrix compared to control composites. This method provides an effective way to introduce chemical linkage between HA filler particles and a polymer matrix. By optimizing the grafting process, a further improvement of the mechanical properties in the composites can be expected.

Covalent bonding of PMMA, PBMA, and poly(HEMA)to hydroxyapatite particles.

In our earlier study, we showed that the surface hydroxyl groups of hydroxyapatite have the ability to react with organic isocyanate groups. In this study, the feasibility of grafting poly(methyl methacrylate) (PMMA), poly(n-butyl methacrylate) (PBMA), and Poly(hydroxyethyl methacrylate) [poly(HEMA)] by using the reaction of isocyanate groups with the hydroxyl groups on the surface of HA was investigated. Double bonds were introduced to the surface of HA via the coupling reaction of isocyanateoethyl methacrylate (ICEM) with HA, or through hexamethylene diisocyanate (HMDI) with hydroxyethyl methacrylate (HEMA) and HA, followed by radical polymerization in MMA, BMA, or HEMA. Infrared spectra indicated the existence of polymers on the surfaces of HA. Thermogravimetric analysis also confirmed the presence of grafted polymers on the surface of HA powder particles (20-26 wt%). The polymers gave typical PMMA, PBMA, or poly(HEMA) infrared spectra, with the exception of amide bands, a result of the coupling reaction of ICEM or HMDI with hydroxy groups of HA or HEMA. Therefore it is concluded that the polymers were chemically bonded to the surface of HA through the isocyanate groups of ICEM or HMDI.