Preparation of an electrodeposited hydroxyapatite coating on titanium substrate suitable for in-vivo applications

In this paper the porous hydroxyapatite coating on Ti implant materials was prepared by the process of electrodeposition, hydrothermal and sinter. The surface morphology, bond strength and thickness of HA coatings were investigated by SEM, AFM, and its biocompatibility was evaluated by cytotoxicity experiments and implant experiments, respectively. Results showed that (1) The HA coatings was 50 μm thickness and adhered on the Ti substrate strongly, which bond strength reached 38MPa. AFM analysis showed that the HA coating was porous structure, in which the mean pore size was 236.5 μm, (2) Cytotoxicity experiments and implant experiments showed that HA-coated Ti implant materials has little cytotoxicity in vitro and little inflammatory reaction in vivo, and there were no statistically disparity between HA-coated Ti implant and titanium implant materials of clinical application (p > 0.05), which demonstrated that HA-coated Ti has a good biocompatibility.     

Powder Metallurgical Near‐Net‐Shape Fabrication of Porous NiTi Shape Memory Alloys for Use as Long‐Term Implants by the Combination of the Metal Injection Molding Process with the Space‐Holder Technique

A new method was developed for producing highly porous NiTi for use as an implant material. The combination of the space‐holder technique with the metal injection molding process allows a net‐shape fabrication of geometrically complex samples and the possibility of mass production for porous NiTi. Further, the porosity can be easily adjusted with respect to pore size, pore shape, and total porosity. The influence of the surface properties of powder metallurgical NiTi on the biocompatibility was first examined using human mesenchymal stem cells (hMSCs). It was found that pre‐alloyed NiTi powders with an average particle size smaller than 45 μm led to the surface properties most suitable for the adhesion and proliferation of hMSCs. For the production of highly porous NiTi, different space‐holder materials were investigated regarding low C‐ and O‐impurity contents and the reproducibility of the process. NaCl was the most promising space‐holder material compared to PMMA and saccharose and was used in subsequent studies. In these studies, the influence of the total porosity on the mechanical properties of NiTi is investigated in detail. As a result, bone‐like mechanical properties were achieved by the choice of Ni‐rich NiTi powder and a space‐holder content of 50 vol% with a particle size fraction of 355–500 μm. Pseudoelasticity of up to 6% was achieved in compression tests at 37 °C as well as a bone‐like loading stiffness of 6.5 GPa, a sufficient plateau stress σ₂₅ of 261 MPa and a value for σ₅₀ of 415 MPa. The first biological tests of the porous NiTi samples produced by this method showed promising results regarding proliferation and ingrowth of mesenchymal stem cells, also in the pores of the implant material.     

Biomorphous porous hydroxyapatite-ceramics from rattan (<Emphasis Type="Italic">Calamus Rotang</Emphasis>)

The three-dimensional, highly oriented pore channel anatomy of native rattan (Calamus rotang) was used as a template to fabricate biomorphous hydroxyapatite (Ca₅(PO₄)₃OH) ceramics designed for bone regeneration scaffolds. A low viscous hydroxyapatite-sol was prepared from triethyl phosphite and calcium nitrate tetrahydrate and repeatedly vacuum infiltrated into the native template. The template was subsequently pyrolysed at 800°C to form a biocarbon replica of the native tissue. Heat treatment at 1,300°C in air atmosphere caused oxidation of the carbon skeleton and sintering of the hydroxyapatite. SEM analysis confirmed detailed replication of rattan anatomy. Porosity of the samples measured by mercury porosimetry showed a multimodal pore size distribution in the range of 300 nm to 300 μm. Phase composition was determined by XRD and FT-IR revealing hydroxyapatite as the dominant phase with minimum fractions of CaO and Ca₃(PO₄)₂. The biomorphous scaffolds with a total porosity of 70–80% obtained a compressive strength of 3–5 MPa in axial direction and 1–2 MPa in radial direction of the pore channel orientation. Bending strength was determined in a coaxial double ring test resulting in a maximum bending strength of ~2 MPa.     

Biodegradable polyurethane composite scaffolds containing Bioglass® for bone tissue engineering

Five types of solid and porous polyurethane composites containing 5–20 wt.% of Bioglass® inclusions were synthesized. Porous structures were fabricated by polymer coagulation combined with the salt-particle leaching method. In-vitro bioactivity tests in simulated body fluid (SBF) were carried out and the marker of bioactivity, e.g. formation of surface hydroxyapatite or calcium phosphate layers upon immersion in SBF, was investigated. The chemical and physical properties of the solid and porous composites before and after immersion in SBF were evaluated using different techniques: Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Dynamic Mechanical Analysis (DMA) and Thermogravimetric Analysis (TGA). Moreover the surface structure and microstructure of the composites was characterised by Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM), respectively. Mercury intrusion porosimetry, SEM and microtomography (μCT) were used to determine pore size distribution and porosity. The fabricated foams exhibited porosity >70% with open pores of 100–400 μm in size and pore walls containing numerous micropores of <10 μm. This pore structure satisfies the requirements for bone tissue engineering applications. The effects of Bioglass® addition on microstructure, mechanical properties and bioactivity of polyurethane scaffolds were evaluated. It was found that composite foams showed a higher storage modulus than neat polyurethane foams. The high bioactivity of composite scaffolds was confirmed by the rapid formation of hydroxyapatite on the foam surfaces upon immersion in SBF.     

Bone ingrowth analysis and interface evaluation of hydroxyapatite coated versus uncoated titanium porous bone implants

Fourteen titanium porous-coated implants with a cylindrical shape (length 22 mm and diameter 5±0.3 mm) were prepared. Bead size was 250–350 m. Seven implants were plasma-sprayed with hydroxyapatite and the other seven remained uncoated. Implants, both hydroxyapatite-coated and uncoated, were randomly selected and press fitted longitudinally into the proximal femoral cancellous bone bilaterally in seven dogs. After 12 weeks the dogs were euthanized and push-out and histomorphometric backscattered electron microscopy studies were carried out. No statistical differences in the mechanical tests were observed. Comparing hydroxyapatite-coated versus uncoated implants, the histomorphometric results showed statistical significance in the percentage of bone (p=0.01); and in bone index, ratio between bone ingrowth and bone ongrowth (p=0.01). The size of the bone implant interface was smaller in the hydroxyapatite applied to spherical bead titanium porous coatings were demonstrated. These morphological and histomorphometric results support the concepts involved with the use of hydroxyapatite as a coating for uncemented porous prosthetic devices.     

Spherical <Emphasis Type="Italic">N</Emphasis>-carboxyethylchitosan/hydroxyapatite nanoparticles prepared by ionic diffusion process in a controlled manner

The nanocomposites containing hydroxyapatite (HA) and biomacromolecules have attracted considerable research interest in implants, tissue scaffolds and drug controlled delivery. In this study, the N-carboxyethylchitosan/hydroxyapatite (NCECS/HA) nanoparticles were prepared by the ionic diffusion process in a controlled manner. The crystallization, particle size, size distribution and aggregation morphology of the NCECS/HA nanocomposites were dependent on the mole ratio of the glucosamine unit in NCECS to the Ca²⁺. Fourier transform-infrared spectroscopic (FTIR) result indicated that there are chemical bonds formed between NCECS and HA. X-ray diffraction (XRD) analysis showed that the crystallization of HA in NCECS matrix was significantly retarded. Transmission electron microscopy (TEM) results revealed that NCECS/HA nanocomposites have the spherical morphology with the diameter ranging from 10 to 40 nm. The NCECS mineralization is driven by the self-assembly of NCECS and HA. These NCECS/HA nanocomposites have potential applications as the carrier for the controlled delivery of growth factors and drugs.     

Preliminary<Emphasis Type="Italic">in vitro</Emphasis> and<Emphasis Type="Italic">in vivo</Emphasis> characterizations of a sol-gel derived bioactive glass-ceramic system

This study investigates quantitatively and qualitatively the sol-gel derived bioactive glass-ceramic system (BGS)—apatite-wollastonite (AW) type granules in the size range of 0.5–1 mm, as an effective graft material for bone augmentation and restoration. Scanning electron micrographs (SEM) of the sintered granules revealed the rough material surface with micropores in the range 10–30 μm. X-ray diffraction (XRD) pattern of the granules revealed the presence of crystalline phases of the hydroxyapatite and wollastonite, and the functional groups of the silicate and phosphates were identified by Fourier transform infrared spectroscopy (FT-IR). Thein vitro cell culture studies with L929 mouse fibroblast cell line showed very few cells adhered on the BGS disc after 24 h. This could be due to the highly reactive surface of the disc concomitant with the crystallization but not due to the cytotoxicity of the material, since the cellular viability (MTT assay) with the material was 80‰ Cytotoxicity and cytocompatibility studies proved that the material was non-toxic and biocompatible. After 12 weeks of implantation of the BGS granules in the tibia bone of New Zealand white rabbits, the granules were found to be well osteointegrated, as observed in the radiographs. Angiogram with barium sulphate and Indian ink after 12 weeks showed the presence of microcapillaries in the vicinity of the implant site implicating high vascularity. Gross observation of the implant site did not show any inflammation or necrosis. SEM of the implanted site after 24 weeks revealed good osteointegration of the material with the newly formed bone and host bone. New bone was also observed within the material, which was degrading. Histological evaluation of the bone healing with the BGS granules in the tibial defect at all time intervals was without inflammation or fibrous tissue encapsulation. After 2 weeks the new bone was observed as a trabeculae network around the granules, and by 6 weeks the defect was completely closed with immature woven bone. By 12 weeks mature woven bone was observed, and new immature woven bone was seen within the cracks of the granules. After 24 weeks the defect was completely healed with lamellar bone and the size of the granules decreased. Histomorphometrically the area percentage of new bone formed was 67.77% after 12 weeks and 63.37% after 24 weeks. Less bone formation after 24 weeks was due to an increased implant surface area contributed by the material degradation and active bone remodeling. The osteostimulative and osteoconductive potential of the BGS granules was established by tetracycline labelling of the mineralizing areas by 2 and 6 weeks. This sol-gel derived BGS granules proved to be bioactive and resorbable which in turn encouraged active bone formation.     

Modell zur Analyse des knöchernen Anwachsverhaltens auf bioaktiv beschichteten Implantatoberflächen

Model to analyse the bone on‐growth on bioactive coated implant surfaces Especially on the field of bone regeneration, transient and permanent implants are an important method of therapy in the Orthopaedic Surgery. In this context, bioactive surfaces on metallic implants provide an improved contact to the surrounding bone. The goal of our study was to establish an in‐vitro test system to evaluate the on‐growth of bone‐derived cells on different surface coatings. Therefore, we invented a special kind of clamps made of commercially‐pure (c‐p) titanium and blasted with hydroxyapatite particles followed by electrochemically coating with calcium phosphate (BONIT®‐HA, BONIT®). Definite pieces of human cancellous bone were attached to these clamps, inserted onto tissue culture plates and cultivated in DMEM for ten days. Finally, the contact area between human cancellous bone and the implant surface was analyzed and the spreading of osteoblast‐like cells evaluated by scanning electron microscopy (SEM). A well‐spread morphology of bone cells was observed on the implant surfaces coated with calcium phosphate (CaP). In comparison the clamps without CaP coatings showed only a marginal growth of bone cells on the clamp surface. The presented newly in‐vitro test setup using titanium clamps coated with bioactive layers attached to human cancellous bone represents a well‐functioning model for qualitative evaluation of bone on‐growth.     

Synthesis of nanocrystalline fluorinated hydroxyapatite by microwave processing and its in vitro dissolution study

Synthetic hydroxyapatite, (Ca₁₀(PO₄)₆(OH)₂, HA), is an important material used for orthopedic and dental implant applications. The biological hydroxyapatite in the human bone and tooth is of nanosize and differs in composition from the stoichiometric HA by the presence of other ions such as carbonate, magnesium, fluoride, etc. Osseointegration is enhanced by using nanocrystalline HA. This stimulates the interest in synthesizing nanocrystalline HA by different routes and among the methods, microwave processing seems to form the fine grain size and uniform characteristic nanocrystalline materials. Fluorinated hydroxyapatite, (FHA, Ca₁₀(PO₄)₆(OH)_2−x F _x ), possesses higher corrosion resistance in biofluids than pure HA and reduces the risk of dental caries. The present work deals with the synthesis of nanocrystalline FHAs by microwave processing. The crystal size and morphology of the nanopowers were examined by X-ray powder diffraction (XRD) and transmission electron microscopy (TEM) methods. The functional groups present in FHA powders were ascertained by Fourier transform infrared spectroscopy (FT-IR) and laser Raman spectroscopy. Since the physiological stability is an important parameter while selecting the material for implantation, the in vitro dissolution studies of FHAs with different fluorine contents were carried out.     

Characterization of porous hydroxyapatite

Hydroxyapatite has been considered for use in the repair of osseous defects for the last 20 years. Recent developments have led to interest in the potential of porous hydroxyapatite as a synthetic bone graft. However, despite considerable activity in this field, regarding assessment of the biological response to such materials, the basic materials characterization is often inadequate. This paper documents the characterization of the chemical composition, mechanical integrity, macro- and microstructure of a porous hydroxyapatite, Endobon ® (E. Merck GmbH), intended for the bone-graft market. Specimens possesed a range of apparent densities from 0.35 to 1.44 g cm^-3. Chemical analysis demonstrated that the natural apatite precursor of Endobon® was not converted to pure hydroxyapatite, but retained many of the ionic substituents found in bone mineral, notably carbonate, sodium and magnesium ions. Investigation of the microstructure illustrated that the struts of the material were not fully dense, but had retained some traces of the network of osteocyte lacunae. Macrostructural analysis demonstrated the complex inter-relationship between the structural features of an open pore structure. Both pore size and connectivity were found to be inversely dependent on apparent density. Furthermore, measurement of pore aspect ratio and orientation demonstrated a relationship between apparent density and the degree of macrostructural anisotropy within the specimens, while, it was also noted that pore connectivity was sensitive to anisotropy. Compression testing demonstrated the effect of apparent density and macrostructural anisotropy on the mechanical properties. An increase in apparent density from 0.38 to 1.25 g cm^-3 resulted in increases in ultimate compressive stress and compressive modulus of 1 to 11 MPa and 0.2 to 3.1 GPa, respectively. Furthermore, anisotropic high density (> 0.9 g cm^-3) specimens were found to possess lower compressive moduli than isotropic specimens with equivalent apparent densities. These results underline the importance of full structural and mechanical characterization of porous ceramic implant materials. ©1999 Kluwer Academic Publishers     

Biocompatible porous ceramics for the cultivation of hematopoietic cells

Porous ceramics made of alumina and hydroxyapatite were created using a protein foaming method. Porosity and pore size distribution were successfully varied by means of chemical modification of the foaming protein Bovine serum albumin (BSA). The effectiveness of the BSA and of its chemical modifications as well as the influence of the dispersing agent were investigated using synchrotron tomography. Resulting porous ceramic materials were used as three-dimensional substrates for the cultivation of human peripheral stem cells. The cells proliferated and differentiated in culture. Five cell lines consistent with human blood cell lines were observed.     

Growth of hydroxyapatite on silica gels in the presence of organic additives: kinetics and mechanism

Synthetic hydroxyapatite was grown on surfaces of silica gels by immersing silica monoliths in a simulated body fluid at 37 °C. The gels were prepared by the sol-gel method and the drying process was controlled by using different additives (mono- and di-ethylene glycol, formamide and glycerin) to obtain large monoliths. The additives affect the growth of the apatite by changing the kinetic constants of chemical reactions on the silica surface. Surface areas and pore size distributions were determined, energy dispersion and FTIR spectra obtained, and scanning electron microscopy performed. The formation of the hydroxyapatite competes with the formation of crystalline calcium carbonate, but the results allow optimization of conditions for the growth of the former as a function of the type and concentration of the additive. For these optimal conditions, the mechanism and the order of the reaction were determined. Received: 15 February 2000 / Reviewed and accepted: 28 June 2000     

Synthesis, characterization and gas sensing property of hydroxyapatite ceramic

Hydroxyapatite (HAp) biomaterial ceramic was synthesized by three different processing routes viz. wet chemical process, microwave irradiation process, and hydrothermal technique. The synthesized ceramic powders were characterized by SEM, XRD, FTIR and XPS techniques. The dielectric measurements were carried out as a function of frequency at room temperature and the preliminary study on CO gas sensing property of hydroxyapatite was investigated. The XRD pattern of the hydroxyapatite biomaterial revealed that hydroxyapatite ceramic has hexagonal structure. The average crystallite size was found to be in the range 31–54 nm. Absorption bands corresponding to phosphate and hydroxyl functional groups, which are characteristic of hydroxyapatite, were confirmed by FTIR. The dielectric constant was found to vary in the range 9–13 at room temperature. Hydroxyapatite can be used as CO gas sensor at an optimum temperature near 125°C. X-ray photoelectron spectroscopic studies showed the Ca/P ratio of 1.63 for the HAp sample prepared by chemical process. The microwave irradiation technique yielded calcium rich HAp whereas calcium deficient HAp was obtained by hydrothermal method.     

A novel bio-inorganic bone implant containing deglued bone, chitosan and gelatin

With the aim of developing an ideal bone graft, a new bone grafting material was developed using deglued bone, chitosan and gelatin. Deglued bone (DGB) which is a by-product of bone glue industries and has the close crystallographic similarities of hydroxyapatite was used as main component in the preparation of bone implant. Chitosan was prepared from the exoskeleton of prawn (Pinaeus indicus, family Crustaceae) which is a by-product of seafood industries. Chitosan gives toughness to the product and do not allow the DGB particles to wither away when the implant is placed in the defect. Gelatin was used as binder for the preparation of DGB-chitosan composite. The DGB, chitosan and DGB-chitosan-gelatin composite, which were prepared in the laboratory, were analysed for their physicochemical properties by infrared spectroscopy, X-ray diffraction and scanning electron microscopy studies.     

Processing Methodologies for Polycaprolactone-Hydroxyapatite Composites: A Review

Biodegradable implants have shown great promise for the repair of bone defects and have been commonly used as bone substitutes, which traditionally would be treated using metallic implants. The need for a second surgery exacerbated by the stress shielding effect caused by an implant has led researchers to consider more effective, synthetic biodegradable graft substitutes. The hierarchical structures commonly designed are inspired by nature in human bones, which consist of minerals such as hydroxyapatite, a form of calcium phosphate and protein fiber. The bone graft bio-substitutes should possess a combination of properties for the purpose of facilitating cell growth and adhesion, a high degree of porosity, which would facilitate the transfer of nutrients and excretion of the waste products, and the scaffold should have high tensile strength and high toughness in order to be consistent with human tissues. Blending of polycaprolactone and hydroxyapatite has demonstrated great potential as bone substitutes. It is essential to identify a standardized processing methodology for the composite, which would result in optimum mechanical property for the biocomposite. In this study, biocomposites made of polycaprolactone (PCL) and hydroxyapatite (HAP) are reviewed for their applications in bone tissue engineering. The processing methodologies are discussed for the purpose of obtaining the porosity and pore size required in an ideal tissue scaffold. The properties of the composite can be varied based on the change in pore size, porosity, and processing methodology. This paper reviews and evaluates the methods to produce the hydroxyapatite-polycaprolactone scaffolds.     

Biocompatible composites of fibrous nanohydroxyapatite embedded in a polydimethylsiloxane

Silicone elastomers have the potential to be a valuable biomaterial due to their mechanical and chemical properties, easy processing, and high gas permeability. Some inherent properties of the pure silicone implant such as high hydrophobicity and low load bearing capacity can be problematic for biomedical applications. The issues were addressed by fabricating hydroxyapatite nanofiber/polydimethylsiloxane nanocomposites. The morphology of nanocomposite structures was visualized by high resolution transmission electron microscopy and field emission scanning electron microscopy. Improved mechanical strength and compliance of the prepared nanocomposite structures were obtained by frequency sweep and creep measurements. Surface hydrophilicity of polydimethylsiloxane was enhanced by hydroxyapatite nanofiber incorporation into the polymer matrix. The cytotoxicity and biocompatibility of the structures were analyzed using breast epithelial cells (MDA MB 231 cell line). These studies showed that the nanocomposite scaffold did not leach any cytotoxic material and showed better cell adhesion and cell proliferation compared to the unfilled elastomer.     

Self-hardening calcium deficient hydroxyapatite/gelatine foams for bone regeneration

In this work gelatine was used as multifunctional additive to obtain injectable self-setting hydroxyapatite/gelatine composite foams for bone regeneration. The foaming and colloidal stabilization properties of gelatine are well known in food and pharmaceutical applications. Solid foams were obtained by foaming liquid gelatine solutions at 50°C, followed by mixing them with a cement powder consisting of alpha tricalcium phosphate. Gelatine addition improved the cohesion and injectability of the cement paste. After setting the foamed paste transformed into a calcium deficient hydroxyapatite. The final porosity, pore interconnectivity and pore size were modulated by modifying the gelatine content in the liquid phase.     

Fracture behavior and biocompatibility evaluation of nylon-infiltrated porous hydroxyapatite

Hybrid hydroxyapatite/polymer composites were prepared by the infiltration of nylon into porous hydroxyapatite. Porous hydroxyapatite (HAp) bodies were prepared from a whisker-like powder with high aspect ratio by pressureless-sintering at various temperatures. Pore characteristics, such as the fraction of open porosity and the pore size distribution, were designed and evaluated by mercury porosimeter. Through the in situ polymerization of -caprolactam, infiltrated into the porous HAp body, a polymeric secondary phase network interpenetrated with the HAp phase was obtained. The obtained hybrid HAp/nylon composites were evaluated with respect to their fracture behavior, i.e., fracture energy, and in vitro bioactivity in simulated body fluid (SBF) in the present paper. These HAp/nylon hybrid composite have a K _IC of 1.65 MPam^1/2 and also a good bioactivity according to the results of SBF immersion tests.     

Antibacterial hydroxyapatite coatings on titanium dental implants

Titanium and its alloys are often used as substrates for dental implants due to their excellent mechanical properties and good biocompatibility. However, their ability to bind to neighboring bone is limited due to the lack of biological activity. At the same time, they show poor antibacterial ability which can easily cause bacterial infection and chronic inflammation, eventually resulting in implant failure. The preparation of composite hydroxyapatite coatings with antibacterial ability can effectively figure out these concerns. In this review, the research status and development trends of antibacterial hydroxyapatite coatings constructed on titanium and its alloys are analyzed and reviewed. This review may provide valuable reference for the preparation and application of high-performance and multi-functional dental implant coatings in the future.     

Histomorphometric study of bone reactions with different hydroxyapatite coating thickness on dental implants in dogs

The purpose of this study was to evaluate the effects of different hydroxyapatite (HA) coating thicknesses on osseointegration after dental implantation in dogs. Six dogs, weighing 10-15 kg, were used in this study. The fixtures, with HA coatings of different thicknesses and made by Dentis, were implanted for 12 weeks in right iliac crestal bone sites and for 6 weeks in left iliac crestal bone sites. Implants in the control group were coated with resorbable blast medium, whereas those in the experimental group were coated with HA of different thicknesses (1, 5, and 15 μm). All dogs were sacrificed at 12 weeks after implant placement, and histological and histomorphometric analyses were performed. The implants were all successful, and a significant difference between the control and experiment groups was observed at 6 weeks, although no significant difference between the control and experimental groups was seen at 12 weeks. These results suggested that HA implant coating may positively affect early osseointegration, but coating thickness had no significant effect on ultimate osseointegration.