J.G.C. Wolke

Calcium phosphate coatings for medical implants

Abstract In surgical disciplines where bone has to be repaired, augmented or improved, bone substitutes are essential. Although bone banks, such as Eurotransplant, are founded to supply such substitutes, natural bone is not always adequate. For example, frequently these so-called bone grafts resorb after implantation (1). Further, they cannot be used for joint and tooth replacement, and recently worries have been raised about the transfer of infectious diseases. Therefore, interest has dramatically increased in the use of synthetic materials for replacement of lost or damaged bone tissue. The generic name of these tissue alternatives is biomaterials. A special class of these biomaterials is composed of metallic devices with coatings to improve bone bonding. These specialized coatings used to improve the metallic implant are the topic of this paper.

Organic-inorganic surface modifications for titanium implant surfaces.

This paper reviews current physicochemical and biochemical coating techniques that are investigated to enhance bone regeneration at the interface of titanium implant materials. By applying coatings onto titanium surfaces that mimic the organic and inorganic components of living bone tissue, a physiological transition between the non-physiological titanium surface and surrounding bone tissue can be established. In this way, the coated titanium implants stimulate bone formation from the implant surface, thereby enhancing early and strong fixation of bone-substituting implants. As such, a continuous transition from bone tissue to implant surface is induced. This review presents an overview of various techniques that can be used to this end, and that are inspired by either inorganic (calcium phosphate) or organic (extracellular matrix components, growth factors, enzymes, etc.) components of natural bone tissue. The combination, however, of both organic and inorganic constituents is expected to result into truly bone-resembling coatings, and as such to a new generation of surface-modified titanium implants with improved functionality and biological efficacy.

Injectable PLGA microsphere/calcium phosphate cements: physical properties and degradation characteristics

Calcium phosphate (CaP) cements show an excellent biocompatibility and often have a high mechanical strength, but in general degrade relatively slow. To increase degradation rates, macropores can be introduced into the cement, e.g., by the inclusion of biodegradable microspheres into the cement. The aim of this research is to develop an injectable PLGA microsphere/CaP cement with sufficient setting/cohesive properties and good mechanical and physical properties. PLGA microspheres were prepared using a water-in-oil-in-water double-emulsion technique. The CaP-cement used was Calcibon®, a commercially available hydroxyapatite-based cement. 10:90 and 20:80 dry wt% PLGA microsphere/CaP cylindrical scaffolds were prepared as well as microporous cement (reference material). Injectability, setting time, cohesive properties and porosity were determined. Also, a 12-week degradation study in PBS (37°C) was performed. Results showed that injectability decreased with an increase in PLGA microsphere content. Initial and final setting time of the PLGA/CaP samples was higher than the microporous sample. Porosity of the different formulations was 40.8% (microporous), 60.2% (10:90) and 69.3% (20:80). The degradation study showed distinct mass loss and a pH decrease of the surrounding medium starting from week 6 with the 10:90 and 20:80 formulations, indicating PLGA erosion. Compression strength of the PLGA microsphere/CaP samples decreased siginificantly in time, the microporous sample remained constant. After 12 weeks both PLGA/CaP samples showed a structure of spherical micropores and had a compressive strength of 12.2 MPa (10:90) and 4.3 MPa (20:80). Signs of cement degradation were also found with the 20:80 formulation. In conclusion, all physical parameters were well within workable ranges with both 10:90 and 20:80 PLGA microsphere/CaP cements. After 12 weeks the PLGA was totally degraded and a highly porous, but strong scaffold remained.

Histomorphometrical and mechanical evaluation of titanium plasma-spray-coated implants placed in the cortical bone of goats

The aim of this study was to investigate the biological and mechanical response of bone to titanium plasma-sprayed implants of different roughnesses. Three types of titanium plasma-spray coating were applied to beam-shaped implants: Ti2, Ti3, and Ti4, with a Ra of 16.5, 21.4, and 37.9 microm, respectively. An Al2O3 grit-blasted implant (Ti-un) with a Ra of 4.7 microm was used as a control. In total, 72 implants were inserted in the tibial cortical bone of nine adult female goats. These implants were evaluated histologically and mechanically 3 months after implantation. At the end of the experiment, of the 72 inserted implants, two implants (one Ti2 and one Ti4) were lost. Histological evaluation of the other retrieved implants revealed a uniform bone reaction for all implants. The unloaded plasma-spray coatings showed no signs of delamination at the implant-coating interface. Occasionally, particles of the Ti4 coating broke free and were found near the implant. Histomorphometry revealed no difference in bone contact for the different implants (P > 0.05). Furthermore, the push-out test showed no significant difference (P > 0.05). Linear regression showed no interaction between the push-out values and the roughness values (r = 0.5). On the basis of these results, it may be concluded that the used surface roughnesses did not lead to differences in bone response or mechanical attachment strength in goat cortical bone.

A histological evaluation of TiO2‐gritblasted and Ca‐P magnetron sputter coated implants placed into the trabecular bone of the goat: part 2

The aim of this study was to investigate the synergetic influence of surface topography and chemical composition of oral implant materials on bone response. For the experiment screw designed implants were used. The implants were grit-blasted with TiO2 particles. The implants were left uncoated (Ti) or provided with three different amorphous/crystalline Ca-P magnetron sputter coatings, resp. 0.1 micron (CaP-0.1), 1 micron (CaP-1) and 4 microns (CaP-4), in thickness. The implants were inserted in the medial femoral condyles of 12 goats. Each femur received 2 implants. After implantation periods of 6 and 12 weeks the implants were retrieved and prepared for histological and histomorphometrical evaluation (bone contact and bone mass). The light microscopy revealed that bone response to CaP-4 and CaP-1 implants was similar. For example, after 12 weeks, screw threads were almost completely covered with bone. In contrast to CaP-0.1 and Ti implants, where bone apposition was less pronounced. Histomorphometry demonstrated that the bone-to-implant contact for the CaP-1 and CaP-4 implants was significantly higher (P < 0.05) than for the CaP-0.1 and Ti implants. This difference existed already after 6 weeks and was even enhanced after 12 weeks. The bone mass measurements revealed that only at 12 weeks CaP-4 implants had significantly more bone contact inside the screw threads than non-coated Ti-implants (P < 0.05). Supported by our findings, we conclude that the additional application of a 1-4 microns thick Ca-P magnetron sputter coating can further improve the healing response to surface roughened oral implants placed into trabecular bone.

Influence of rapid heating with infrared radiation on RF magnetron-sputtered calcium phosphate coatings

This study evaluated the effect of rapid heating with infrared radiation on the physico-chemical and morphological properties of radio frequent (RF) magnetron-sputtered calcium phosphate (Ca-P) coatings. About 2.5 microm thick Ca-P coatings were deposited on titanium disks and cylinders. These specimens were left untreated or were heat treated by infrared radiation at 300, 400, 500, 600, and 700 degrees C for 4, 7, 11, 17, and 24 s. Subsequently, the specimens were immersed in simulated body fluid (SBF) for 1 day, 1 week, and 5 weeks. X-ray diffraction measurements showed that heating at 500 degrees C or higher resulted in an increase of coating crystallinity. In addition, FT-IR measurements revealed the appearance of OH peaks in the spectra of samples treated at 500-700 degrees C. Electron probe microanalysis showed that after 5 weeks of immersion about 40-50% of the coatings heat treated at 500 and 600 degrees C was maintained. The coatings heat treated at 700 degrees C showed no dissolution at all. On the other hand, as-coated and 300 degrees C treated films were dissolved within 1 day. Scanning electron microscopy of the samples showed that directly after heat treatment no apparent cracks were present in the coatings. On the basis of these findings, we conclude that rapid heating with infrared radiation around 600 degrees C is the best heat treatment for RF magnetron-sputtered coatings.

Bone response to fast-degrading, injectable calcium phosphate cements containing PLGA microparticles

Apatitic calcium phosphate cements (CPC) are frequently used to fill bone defects due to their favourable clinical handling and excellent bone response, but their lack of degradability inhibits complete bone regeneration. In order to render these injectable CaP cements biodegradable, hollow microspheres made of poly (D,L-lactic-co-glycolic) acid (PLGA) have been previously used as porogen since these microspheres were shown to be able to induce macroporosity upon degradation as well as to accelerate CPC degradation by release of acid degradation products. Recently, the capacity of PLGA microspheres to form porosity in situ in injectable CPCs was optimized by investigating the influence of PLGA characteristics such as microsphere morphology (dense vs. hollow) and end-group functionalization (acid terminated vs. end-capped) on acid production and corresponding porosity formation in vitro. The current study has investigated the in vivo bone response to CPCs containing two types of microspheres (hollow and dense) made of PLGA with two different end-group functionalizations (end capped and acid terminated). Microspheres were embedded in CPC and injected in the distal femoral condyle of New Zealand White Rabbits for 6 and 12 weeks. Histological results confirmed the excellent biocompatibility and osteoconductivity of all tested materials. Composites containing acid terminated PLGA microspheres displayed considerable porosity and concomitant bone ingrowth after 6 weeks, whereas end capped microspheres only revealed open porosity after 12 weeks of implantation. In addition, it was found that dense PLGA microspheres induced significantly more CPC degradation and bone tissue formation compared to hollow PLGA microspheres. In conclusion, it was shown that PLGA microspheres have a strong capacity to induce fast degradation of injectable CPC and concomitant replacement by bone tissue by controlled release of acid polymeric degradation products without compromising the excellent biocompatibility and osteoconductivity of the CPC matrix.

The osteogenic effect of electrosprayed nanoscale collagen/calcium phosphate coatings on titanium.

For orthopedic and dental implants, the ultimate goal is to obtain a life-long secure anchoring of the implant in the native surrounding bone. To this end, nanoscale calcium phosphate (CaP) and collagen-CaP (col-CaP) composite coatings have been successfully deposited using the electrospray deposition (ESD) technique. In order to study to what extent the thickness of these coatings can be reduced without losing coating osteogenic properties, we have characterized the mechanical and biological coating properties using tape tests (ASTM D-3359) and in vitro cell culture experiments, respectively. Co-deposition of collagen significantly improved coating adhesive and cohesive strength, resulting in a remarkably high coating retention of up to 97% for coating thicknesses below 100 nm. In vitro cell culture experiments showed that electrosprayed CaP and col-CaP composite coatings enhanced osteoblast differentiation, leading to improved mineral deposition. This effect was most pronounced upon co-deposition of collagen with CaP, and these coatings displayed osteogenic effects even for a coating thickness of below 100 nm.

Soft-tissue response to injectable calcium phosphate cements.

In this study, the soft tissue reaction to two newly developed injectable calcium phosphate bone cements (cement D and W) was evaluated after implantation in the back of goats. For one of the cements (cement D) the tissue reaction was also investigated after varying the concentration of accelerator Na(2)HPO(4) in the cement liquid (resulting in cement D1 and D2). Eight healthy mature female Saanen goats were used. The cement was applied 10min after mixing while it was still moldable and plastic. The material was given a standardized cylindrical shape. Thirty-two implants of each cement formulation were inserted and left in place for 1, 2, 4, and 8weeks. At the end of the study, eight specimens of each material and healing period were available for further analysis. Two specimens were used for X-ray diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR) and six specimens were used for light microscopical evaluation. XRD and FTIR showed that the cements did set as microcrystalline carbonate apatite with the disappearance of monetite from the cements during implantation. Histological analysis showed that after 8weeks of implantation around all materials a thin soft-tissue capsule was formed (thickness ranging from 5 to 15 cell layers) with almost complete absence of inflammatory cells. Only in some specimens a slightly higher inflammatory reaction was observed. This was due to cement surface defects and a zone of dispersed particles near the cement-soft tissue interface. There was almost no resorption of the material after 8 weeks of implantation. In a few 4 and 8weeks samples, small areas of calcification were found in the fibrous capsule surrounding the implants. On the basis of our observations, we conclude that the tested cements were biocompatible and can be used next to soft tissue.

Introduction of gelatin microspheres into an injectable calcium phosphate cement.

For tissue engineered bone constructs, calcium phosphate cement (CPC) has a high potential as scaffold material because of its biocompatibility and osteoconductivity. However, in vivo resorption and tissue ingrowth is slow. To address these issues, microspheres can be incorporated into the cement, which will create macroporosity after in situ degradation. The goal of this study was to investigate the handling properties and degradation characteristics of CPC containing gelatin microspheres. Setting time and injectability were determined and an in vitro degradation study was performed. Samples were assayed on mass, compression strength, E-modulus, and morphology. A supplementary degradation test with gelatin microspheres was performed to investigate the influence of physical conditions inside the cement on microsphere stability. The gelatin microsphere CPCs were easy to inject and showed initial setting times of less than 3 min. After 12-weeks in vitro degradation no increase in macroporosity was observed, which was supported by the small mass loss and stabilizing mechanical strength. Even a clear densification of the composite was observed. Explanations for the lack of macroporosity were recrystallization of the cement onto or inside the gelatin spheres and a delayed degradation of gelatin microspheres inside the scaffold. The supplementary degradation test showed that the pH is a factor in the delayed gelatin microsphere degradation. Also differences in degradation rate between types of gelatin were observed. Overall, the introduction of gelatin microspheres into CPC renders composites with good handling properties, though the degradation characteristics should be further investigated to generate a macroporous scaffold.