[Attaching single- and multi-unit fixed dental prostheses]. / Bevestiging van kronen en bruggen.

A single- or multi-unit fixed dental prosthesis can be attached to the abutment teeth through mechanical retention and gap sealing or by adhesion. For sealing the gap, water-soluble cements are appropriate, such as zinc phosphate, polycarboxylate, and (resin-modified) glasionomer cement. Attachment through adhesion can be performed with composite cement. If the hard tooth tissue is prepared adequately, composite cement provides sufficient adhesion, but self-adhesive composite cement is now also available. For the adhesion of the composite cement to the restorative materials of the single- or multi-unit fixed dental prosthesis, surface sandblasting, silanizing, and tin coating and the application of a metal primer or chemically active composite are available. Cementing a single- or multi-unit dental prosthesis involves 3 phases: 1. Cleansing the single- or multi-unit dental prosthesis and the abutment tooth/teeth; 2. Preparing the hard tooth tissue, mixing the cement and placing the single- or multi-unit dental prosthesis; 3. Removing the excess cement.

Ceramic composites as matrices and scaffolds for drug delivery in tissue engineering.

Ceramic composites and scaffolds are popular implant materials in the field of dentistry, orthopedics and plastic surgery. For bone tissue engineering especially CaP-ceramics or cements and bioactive glass are suitable implant materials due to their osteoconductive properties. In this review the applicability of these ceramics but also of ceramic/polymer composites for bone tissue engineering is discussed, and in particular their use as drug delivery systems. Overall, the high density and slow biodegradability of ceramics is not beneficial for tissue engineering purposes. To address these issues, macroporosity can be introduced often in combination with osteoinductive growth factors and cells. Ceramics are good carriers for drugs, in which release patterns are strongly dependent on the chemical consistency of the ceramic, type of drug and drug loading. Biodegradable polymers like polylactic acid, gelatin or chitosan are used as matrices for ceramic particles or as adjuvant to calcium phosphate cements. The use of these polymers can introduce a tailored biodegradation/drug release to the ceramic material.

The in vitro behavior of as-prepared and pre-immersed RF-sputtered calcium phosphate thin films in a rat bone marrow cell model.

In this paper we focus on the behavior of radio frequency (RF)-sputtered calcium phosphate (CaP) thin films in a rat bone marrow (RBM) cell model. Two issues are addressed. Firstly, we benchmarked the in vitro cell behavior of these CaP coatings by comparing their proliferation, differentiation and mineralization behavior and the structure of the formed interface to similar coatings of alumina and titania. We found that the CaP coatings showed reduced proliferation, enhanced early differentiation and enhanced activity of mature osteoblasts compared to the alumina coatings. Enhanced production of mineralized extracellular matrix (ECM) was seen for both CaP and titania. Two types of CaP precipitates could be observed, one directly bonded CaP layer at the coating interface and one of globular accretions associated with the ECM. The directly bonded layer was not observed on the alumina coatings. Further, no thin film effects were found. Secondly, the effect of pre-immersion of the CaP coatings in SBF2 was explored. We found that the early formation of a directly bonded CaP layer is obstructed by the absence of CaP nuclei. After approximately 8 days, cell activity induces the nucleation of CaP crystals on both the surface and the ECM, and growth is enhanced. By initially providing these coatings with CaP crystals, growth of the directly bonded CaP layer is immediate. Hence, the formation of the interfacial CaP layer and the matrix-associated CaP accretions can effectively be decoupled.

Subcutaneous evaluation of RF magnetron-sputtered calcium pyrophosphate and hydroxylapatite-coated Ti implants.

The in vivo behavior of infrared-heated, RF magnetron-sputtered hydroxylapatite (HA) and calcium pyrophosphate (DCPP) coated titanium discs was investigated. The discs were implanted subcutaneously in the back of six goats for 2, 4, 8 and 12 weeks. At the end of the study, coated discs were removed and examined on their physicochemical properties by X-ray diffraction (XRD) and scanning electron microscopy (SEM), including energy dispersive spectroscopy (EDS). Also, implants were prepared for light microscopical evaluation of the tissue response. The results showed that heat-treated HA coatings showed a stable behavior, i.e. no changes in the XRD pattern occurred during implantation. Also, no dissolution of the coating was observed by SEM. EDS revealed that the Ca/P ratio of the HA coatings remained stable during implantation. In contrast, heat-treated DCPP coatings showed a compositional change into apatite and tricalcium phosphate (TCP) during implantation. This was confirmed by the SEM and EDS analysis. The Ca/P ratio of the DCPP coatings changed from 0.8 to 1.52 during implantation. Finally, histology showed that both heat-treated HA and DCPP coatings showed no adverse tissue response, as characterized by the presence of thin, dense fibrous tissue capsule. Consequently, it can be concluded that 2 mum thick heat-treated, RF magnetron-sputtered HA and DCPP coatings are of sufficient thickness to withstand dissolution during 12 weeks of implantation in a subcutaneous location in goats. In addition, both coatings showed a biocompatible tissue behavior. Further, heat-treated DCPP coatings revealed a gradual compositional change into apatite and TCP.

Preparation and characterization of RF magnetron sputtered calcium pyrophosphate coatings.

CaP ceramic has been widely used as coating on metals in orthopedics and oral dentistry. Variations in CaP composition can lead to different dissolution/precipitation behavior and may also affect the bone response. In the present study calcium pyrophosphate and hydroxylapatite coatings were successfully prepared by RF magnetron sputtering deposition. The phase composition, morphological properties, and the dissolution in SBF were characterized by using XRD, FTIR, EDS, SEM, and spectrophotometry. The results showed that all the sputtered coatings were amorphous and changed into a crystal structure after IR-radiation. The temperature for the crystallization of the amorphous coatings is lower for the hydroxylapatite coating (550 degrees C), compared to the calcium pyrophosphate coating (650 degrees C). All sputtered amorphous coatings were instable in SBF and dissolved partially within 4 wks of incubation. The heat-treated coatings appeared to be stable after incubation. These results showed that magnetron sputtering of calcium pyrophosphate coating is a promising method for forming a biocompatible ceramic coating.

Growth behavior of rat bone marrow cells on RF magnetron sputtered hydroxyapatite and dicalcium pyrophosphate coatings.

The aim of this study was to evaluate the osteogenic properties of magnetron sputtered dicalcium pyrophaosphate (DCPP) and hydroxylapatite (HA) coatings. Therefore, DCPP and HA coatings were deposited on grit-blasted titanium discs. The substrates were used as-prepared or received an additional heat treatment which changed the amorphous coating structure to a crystalline structure. Subsequently, rat bone marrow stromal cells were cultured for 1-24 days on the various substrates. DNA and alkaline phosphatase activity was determined after 1, 3, 5, 8, and 12 days of incubation. Osteocalcin expression was evaluated after 8, 12, 16, and 24 days of incubation. Scanning electron microscopical analysis of cell morphology and coating characteristics was done after 8 and 16 days of incubation. All assays were done in duplicate and in each assay all specimens were present in fourfold. Results demonstrated that the cells did not proliferate and differentiate on all amorphous coatings. SEM revealed that the amorphous coatings showed significant dissolution. On the crystalline DCPP and HA coatings an increase in DNA and alkaline phosphatase activity was seen starting at day 8 of incubation. Osteocalcin expression on the crystalline coatings started to increase at day 16 of incubation. SEM showed that the growth and differentiation of the cells was associated with extensive collagen fiber formation and surface mineralization in the form of globular accretions. Further, statistical testing revealed that proliferation and differentiation of the rat bone marrow stromal cells started significantly earlier on the crystalline HA coatings than that on the crystalline DCPP coatings. These results demonstrate that the rat bone marrow stromal cells proliferated and differentiated only on crystalline magnetron sputtered DCPP as well as HA coatings, which warrants the further in vivo analysis of the bone healing supporting properties of these coatings.

The influence of the crystallinity of electrostatic spray deposition-derived coatings on osteoblast-like cell behavior, in vitro.

This article describes the influence of the crystallinity of carbonate apatite (CA) coatings on osteoblast-like cell behavior. Porous CA coatings were produced with electrostatic spray deposition (ESD), and subsequently, received heat treatments of 400, 500, or 700 degrees C to induce various coating crystallinities. As a result, an amorphous calcium phosphate (ACP), a crystalline CA (CCA), and a crystalline carbonated hydroxyapatite (CHA) structure were formed, respectively. Uncoated titanium substrates served as the control group. After seeding rat osteoblast-like cells, the initial cell attachment was similar between the groups, and approached 100% after 6 h. Between the various coatings, no differences were observed for proliferation, differentiation, or mineralization. However, proliferation of the osteoblast-like cells was lower on all coated substrates after longer culture periods, compared to the uncoated substrates, while at the same time differentiation was stimulated. Furthermore, after 8 and 16 days of incubation, scanning electron microscopy showed more signs of mineralization on coated substrates, compared to the uncoated substrates. In conclusion, porous ESD-derived CA coatings have a positive effect on the in vitro differentiation of osteoblast-like cells, compared to uncoated, as-machined titanium. However, this effect is not further enhanced by the degree of crystallinity of the ESD-derived CA coatings.

Mechanical properties of porous, electrosprayed calcium phosphate coatings.

Mechanical properties of calcium phosphate coatings (CaP), deposited using the electrostatic spray deposition (ESD) technique, have been characterized using a range of analytical techniques, including tensile testing (ASTM C633), fatigue testing (ASTM E855), and scratch testing using blunt and sharp scratch styli. Moreover, a simple explantation procedure was successfully introduced using ESD-coated, threaded dental implants to characterize the mechanical performance of CaP coatings qualitatively under conditions that mimic clinical situations as close as possible. Generally, all analysis techniques revealed that ESD coatings need to be crystallized in order to ensure interfacial adhesion to the substrate and sufficient mechanical strength of the superficial reticular structure. Crystalline carbonated hydroxyapatite coatings (CHA, heat-treated at 700 degrees C) were resistant to fatigue as well as to plastic ploughing deformation by means of various scratch styli, and the fragile surface structure of ESD coatings was maintained to a large extent after unscrewing CHA-coated dental implants from femoral condyles of goat cadavers. From these experiments, it was concluded that interfacial adhesion of crystalline CHA ESD coatings to the titanium substrate was sufficient, but that mechanical strength of the superficial architecture of ESD coatings need to be optimized for applications where high shear and compressive stresses are imposed onto the rather fragile coating surface of reticular ESD morphologies.

Closing capacity of cranial bone defects using porous calcium phosphate cement implants in a rabbit animal model.

Calcium phosphate (Ca-P) cement is a well established material for bone repair. The bone biological properties of Ca-P cement can even be further improved by creating porosity in the material. The current study aimed on the evaluation of the osteoconductive behavior of porous Ca-P cement. Therefore, circular defects (6, 9, and 15 mm in diameter) were created in the cranium of 3 months old rabbits and filled with porous Ca-P cement implants. The total porosity of implants was calculated to be 71, 74 and 74% respectively and the average pore diameter was 150 microm. In addition, empty control defects were prepared. After 12 weeks implantation time the animals were sacrificed and radiographic, histological, and histomorphometrical evaluation was performed. The Critical Size Defect (CSD) of this species at this location for an implantation time of 12 weeks was confirmed to be 15 mm. Bone was observed to be present over and through almost all porous Ca-P cement implants. Only, in one out of eight animals with a 15 mm implant complete bone bridging of the defect did not occur. The size of the defect was found not to affect the total percentage of bone formation in the cement; (17 +/- 7)%, (18 +/- 6)% and (17 +/- 3)% for respectively 6, 9, and 15 mm diameter implants. We concluded that porous Ca-P cement is an excellent osteoconductive material in non weight bearing situations and complete bridging of a critical sized skull defect occurs in this rabbit model after 12 weeks of implantation.

Differentiation ability of rat postnatal dental pulp cells in vitro.

The current rapid progression in stem cell research has enhanced our knowledge of dental tissue regeneration. In this study, rat dental pulp cells were isolated and their differentiation ability was evaluated. First, dental pulp cells were obtained from maxillary incisors of male Wistar rats. Immunochemistry by stem cell marker STRO-1 proved the existence of stem cells or progenitors in the isolated cell population. The dissociated cells were then cultured both on smooth surfaces and on three-dimensional (3-D) scaffold materials in medium supplemented with beta-glycerophosphate, dexamethasone, and L-ascorbic acid. Cultures were analyzed by light and scanning electron microscopy and, on proliferation, alkaline phosphatase activity and calcium content were determined and the polymerase chain reaction was performed for dentin sialophosphoprotein, osteocalcin, and collagen type I. These cells showed the ability to differentiate into odontoblast-like cells and produced calcified nodules, which had components similar to dentin. In addition, we found that the "odontogenic" properties of the isolated cells were supported by three-dimensional calcium phosphate and titanium scaffolds equally well.

Osteoclastic resorption of calcium phosphate coatings applied with electrostatic spray deposition (ESD), in vitro.

Calcium phosphate (CaP) coatings have been applied on titanium implants to improve the bioactivity in order to favor the initial bone healing response. Recently, a new technique has been developed to apply CaP coatings: electrostatic spray deposition (ESD). Although ESD-derived coatings have several benefits, it is not known whether they are degradable. This study was designed to examine the cell-mediated degradation of two ESD-derived coatings with different chemical compositions, that is, beta-tricalcium phosphate (beta-TCP) and carbonate apatite (CA). First, coatings were deposited and analyzed physiochemically. Subsequently, rat bone marrow-derived osteoclastlike cells were seeded on the coatings, and analyzed with osteoclast-specific markers, scanning electron microscopy, and transmission electron microscopy. Results showed that both coatings exhibited porous morphologies, with an average pore size of less than 1 microm (beta-TCP), or larger than 1 microm (CA). After heat treatment, both coatings were crystalline in structure. The Ca/P ratios were 1.4 to 1.5 for the beta-TCP coating, and 1.8 to 2.0 for the CA coating. After 8 and 12 days of culture, multinucleated osteoclastlike cells were observed on both coatings. The osteoclast phenotype was confirmed by tartrate resistant acid phosphatase (TRAP) staining, and immunostaining against the calcitonin receptor. Using scanning electron microscopy, numerous resorption lacunae were observed in both coatings. Finally, transmission electron microscopy of TRAP-positive cells confirmed the osteoclastlike aspect of the cells revealing multiple nuclei and a ruffled border. In conclusion, CaP coatings produced with the ESD process can be degraded by osteoclasts.

Microstructure and crystallinity in hydroxyapatite coatings.

Plasma spraying was used to produce hydroxyapatite (HA) coatings on metal. The microstructure of these coatings, sprayed with two powder particle size distributions, was examined by scanning electron microscopy (SEM) and showed scattered HA particles with completely melted and unmelted or partially melted cores. Röntgen diffraction shows that the crystallinity increases after a vacuum heat treatment at 600 degrees C. SEM reveals that the amorphous phase recrystallized and new crystalline grains were formed at the surface of the crystalline cores.

The effect of titanium plasma-sprayed implants on trabecular bone healing in the goat.

The bone response to different titanium plasma-sprayed implants was evaluated in a goat model. Therefore, beam-shaped implants were installed into the trabecular femoral condyles of 10 goats. These implants were provided with three different titanium plasma-sprayed coatings (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 control. After an implantation period of 3 months, the implants were evaluated histologically and histomorphometrically. Only one implant (Ti3) was not recovered after the evaluation period. Light microscopy showed a limited amount of bone for the various implants. Most of the implants showed a different degree of fibrous tissue alternating with direct bone contact. Complete fibrous encapsulation of the implants was observed in some of the sections. No signs of delamination of the plasma-sprayed coating was visible. No significant difference in bone contact were measured between the different types of implants (P > 0.05). Histomorphometrical analysis revealed significantly higher bone mass close to the implant (0-500 microm) for the Ti3, Ti4 and Ti-un implants placed in the medial femoral condyle and the Ti4 implants placed in the lateral condyle. At distance (500-1500 microm), no difference in bone mass measurements between the different implants was observed (P > 0.05).

Influence of annealing temperature on RF magnetron sputtered calcium phosphate coatings.

The effect of different annealing temperatures on the characteristics of thin calcium phosphate coatings fabricated by radiofrequency magnetron sputtering was studied. Annealing of the as-sputtered films was necessary to change the amorphous coating to a crystalline coating. The films were annealed for 2 and 4 h at 400, 600, 800, 1000 and 1200 degrees C under dry argon or argon and water vapour flow. After annealing, the structure and the chemical composition of these films were characterized with incident light microscopy, Rutherford backscattering spectrometry (RBS), X-ray diffraction (XRD), and Fourier transform infrared absorption spectrometry (FTIR). Incident light microscopy showed cracks in the coatings annealed at a higher temperature than 400 degrees C. RBS revealed that the as-sputtered coatings had a high Ca/P ratio which decreased with increasing annealing temperature. After annealing at a temperature of 600 degrees C or more the XRD showed crystalline hydroxyapatite (HA) coatings. However, the second phase, present in the coatings, changed from tetra-calcium phosphate to calcium oxide to beta-tri-calcium phosphate with increasing annealing temperature. FTIR measurements showed the existence of OH- and PO- bonds in all coatings, although the PO- bonds varied for different annealed coatings, from the PO- bonds due to HA to PO- bonds due to other calcium phosphates. From the results of this study we suggest that 600 degrees C is probably the best annealing temperature to obtain a better characterization and understanding of the coating.

Formation and characteristics of the apatite layer on plasma-sprayed hydroxyapatite coatings in simulated body fluid.

Plasma-sprayed hydroxyapatite (HA) coatings were incubated in simulated body fluids (SBFs) for different periods of time to investigate the nucleation and growth of apatite on their surface. The layer that formed was recognized as having similarities to bone apatite because it is poorly crystallized, non-stoichiometric or calcium deficient, and contains carbonate and magnesium. Scanning electron microscopy (SEM) and infrared spectroscopy (IR) were employed to investigate the morphological changes of the coating surface and the structure of the grown layer respectively. In the first few hours, calcium and phosphate ions dissolved from the coatings so as to increase their local supersaturation to a higher degree, thereafter followed by the nucleation and growth of apatite. The nucleation occurred firstly on the recessed regions, inside pores and cracks where the higher supersaturation was readily maintained. Only after 24 h incubation was a complete layer formed on the surface of the coating. There is no obvious interface between the grown layer and the underlying coating. Heat treatment in the air made the apatite transform into biphasic calcium phosphate of HA and tricalcium phosphate, with a blue colour because of trace manganese ions. The heat-treated HA coating showed no dissolution by SEM observation. This resulted in no precipitation on the surface. When SBF was used with two-fold higher ion concentrations, the apatite layer formed slowly in 72 h without dissolution of the coating surface. This may mean that the microenvironment with a sufficiently high degree of supersaturation of calcium and phosphate ions is crucial for apatite to nucleate and grow in SBF, while the HA crystalline structure is not critical in the nucleation process, as expected.

A new method to produce macropores in calcium phosphate cements.

A new way to create macropores in calcium phosphate cements has been developed. The method consists in adding NaHCO3 to the starting cement powder (Biocement D) and using two different liquids: first a basic liquid to form the paste and later an acid liquid to obtain CO2 bubbles. Mercury intrusion measurements showed a dramatic increase both in macropores with an average size of 100 m and in the total porosity (even higher than 50% with respect to the Biocement D). This method does not change in any significant way the final reaction products of the starting material after being soaked 3 days in Ringer solution. Only, due to the increase of the porosity. the compressive strength of the porous cement decreases significantly.

Stability of radiofrequency magnetron sputtered calcium phosphate coatings under cyclically loaded conditions.

The stability of radiofrequency (RF) magnetron sputtered calcium phosphate was studied under cyclically loaded conditions. The coatings were deposited on titanium bars and tested in either dry or wet conditions X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray (EDX) analysis and Fourier transform infrared (FTIR) spectroscopy were used to characterize the as-sputtered and tested coatings. XRD demonstrated that the amorphous structure after annealing at 650 C changed into a crystalline apatite structure. The residual stresses were determined by the XRD cos 2 i/i method. These residual film stresses were influenced by the coating conditions and the crystalline sputtered coating showed the presence of compressive stresses. SEM demonstrated that, after cyclic loading conditions in air, the crystalline sputter-coated Ti-6A1-4V bars showed a partial coating loss. Furthermore, in wet conditions (simulated body fluid) only the heat-treated sputter-coated bars appeared to be stable. On the other hand, the amorphous coating only showed signs of delamination in the more highly stressed regions, while in the less stressed regions a Ca-P precipitate was formed. On the basis of these results we conclude that calcium phosphate coatings subjected to cyclic loading conditions show an important difference in fatigue behaviour when tested in either dry or wet conditions.

Long-term in vivo study of plasma-sprayed coatings on titanium alloys of tetracalcium phosphate, hydroxyapatite and alpha-tricalcium phosphate.

In order to study the interaction of calcium phosphate coatings with bone tissue, coated titanium plugs of standard size were implanted in dog femora. The bone bonding and bone formation of hydroxyapatite, alpha-tricalcium phosphate (alpha-TCP) and tetracalcium phosphate plasma-sprayed coatings were evaluated by mechanical push-out tests and histological observations after 3, 5, 15 and 28 months of implantation. During this time all coating types degraded. alpha-TCP showed the most significant degradation after 3 months of implantation. Hydroxyapatite and tetracalcium phosphate showed significant signs of degradation after about 5 months of implantation. All coatings showed a small increase in bone bonding after 5 months of implantation. In general, all types of implants showed similar bone response, some bone contact and several remodelling lacunae along the surfaces after long-term implantation.

Studies of the solubility of different calcium phosphate ceramic particles in vitro.

In vitro solubility tests of hydroxyapatite, tetracalcium phosphate or tricalcium phosphate particles were performed in lactate, citrate, Gomoris or Michaelis buffer with pH 6.2 or 7.2 and in aqua destillata. The results showed that in general the solubility decreased in the order tetracalcium phosphate greater than tricalcium phosphate greater than hydroxyapatite, except for lactate or citrate buffer where the solubility order was tetracalcium phosphate = tricalcium phosphate greater than hydroxyapatite. The influence of the specific buffer used is much larger than either pH or specific calcium phosphate salt tested. The pH stability of lactate buffer and aqua destillata is very low, the other buffer solvents had a rather stable pH value.

Initial deposition of calcium phosphate ceramic on polyethylene and polydimethylsiloxane by rf magnetron sputtering deposition: the interface chemistry.

Calcium phosphate (CaP) coatings are well known for their bioactive nature. CaP coated polymeric materials can be used as implant material. For this, a strong adhesion between the coating and substrate is necessary. Because the chemical structure of the interface plays an important role in the coating adhesion, we studied the interface between CaP and the polymers polyethylene (PE) and polydimethylsiloxane (PDMS/silicone rubber). Both untreated and plasma pretreated polymers were used. On PE, a low Ca/P ratio nearby the interface and a high amount of C-O bonds were found on both untreated and plasma pretreated PE. This is the result of phosphate-like groups that are able to bind to the carbon of the PE. PDMS reacts towards the plasma pretreatment by losing CH(3) side groups. Compared to PE, a low amount of C-O bonds is found nearby the interface. Besides, a low Ca/P ratio is found nearby the interface. This is the result of phosphate groups that connect to Si atoms of the PDMS, thereby replacing the CH(3) side groups. The bombardment by negatively charged oxygen ions that are accelerated from the target during the deposition process makes the chemical interaction between the coating and the substrates possible.