Immediate implant placement using injectable calcium phosphate hydraulic cement in dogs.

Techniques allowing implant placement in extraction sockets require either high diameter implants in surgically enlarged sockets or grafting and/or regenerative procedures around implants after their primary surgical stabilization. This study aimed to evaluate the ability of calcium phosphate hydraulic cement (CPHC) to immobilize commercially available titanium implants in extraction sockets. CPHC was used in seven fresh dog extraction sockets in conjunction with ITI TPS implants. Three extraction sockets without CPHC were used as controls. Initial implant stability was measured after 10 min hardening with periostest. The dogs were sacrificed after 9 months. Non-decalcified specimens were prepared for histologic and histomor-phometric examination. The surface percentage of implant-to-mineralized bone contact and bone density was calculated for each specimen. The periotest values were significantly different for implants stabilized with CPHC than for the controls, and simi-lar to values reported for osteointegrated implants. New alveolar bone was formed in intimate contact with titanium. In two cases, non-resorbed CPHC residues were observed closely bound to the implant. This study clearly demonstrates that CPHC cement is suitable for immediate implant immobilization in extraction sockets.

Influence of polymeric additives on the biological properties of brushite cements: an experimental study in rabbit.

The resorbability and ability of calcium phosphate hydraulic cements to promote new bone formation was investigated in vivo. The effects of two hydrosoluble polymeric additives (hyaluronic acid, and xanthan gum,) on the biological response of two brushite cement formulations (BHC-A vs BHC-B) was investigated. The brushite cements differed in P/Ca (0.71 vs 0.98) and S/Ca (0.10 vs 0.005) atomic ratios and by the presence of calcium sulfate hemihydrate in BHC-A. Polymer-free cements were used as controls. Cement specimens were injected in cylindrical bone defects manually drilled in the distal condyle of rabbit femora. The implants were harvested at 12 and 24 weeks after implantation and subjected to quantitative histomorphometry. The study showed a significantly lower resorption rate for cement BHC-A, which induces the formation of well-mineralized bone in close apposition to the residual material. In contrast, cement BHC-B showed a significant increase of bone formation period and the formation of a thick layer of unmineralized osteoid tissue at the bone/residual cement interface. The presence of xanthan gum made the biological response even worse, particularly in the case of cement BHC-B. The presence of hyaluronic acid has little effect, except for a slight decrease in initial resorption rate, in the case of cement BHC-A.

Injectable calcium phosphate hydraulic cement (CPHC) used for periodontal tissue regeneration: A study of a dog model.

Injectable calcium phosphate hydraulic cement (CPHC) is a new bone substitute family. This study aimed to evaluate the use of CPHC in surgical periodontitis-simulating defects in a dog model. CPHC was obtained by adding powder mixtures of different calcium phosphates with different solubility. Alveolar bone was removed by drilling over the mesial and distal roots of the 2nd mandibular premolar in six dogs. The defects were randomly selected, three were untreated and six treated. The defects had a depth of 6 mm and a width of 3 mm. The animals were sacrificed after 9 months and samples prepared, with no decalcification, for histological evaluation. Seventy-nine percent of the root was covered by bone in the experimental defects, compared to 41% of the root for the control defects. Bone height was significantly higher for the experimental defects (4.9 +/- 0.9 mm) than for the control defects (1.4 +/- 0.5 mm). After 9 months, 97 +/- 6% of the CPHC was degradated and replaced by bone. This study proves the interest of this cement because of the particularly high level of periodontal bone regeneration. The ability of the cement to be easily injected and shaped in bone defects and the immediate immobilization of the teeth after hardening is notable. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 186-93).

Bone colonization of beta-TCP granules incorporated in brushite cements.

Injectable calcium phosphate hydraulic cements are known to have a high clinical potential in bone reconstruction for mini-invasive orthopaedic surgery, interventional radiology, and rheumatology. Previous in vivo experiments in rabbit have shown that the presence of beta-TCP granules in injectable bone cement help maintain the transient biomechanical function of the implanted bone and promote the formation of good-quality new bone. Histomorphometric analysis of two brushite hydraulic cement (BHC) mixtures selected from previous results (referred to in this work as BHC-A and BHC-B) was performed at three postoperative delays (0, 12, and 24 weeks): histomorphometric analysis of bone colonization within beta-TCP shows that, just before implantation, the beta-TCP granule area is significantly higher in BHC-B; the residual granule area decreases steadily over time in BHC-A, whereas it goes through a maximum of 30% at 12 weeks in BHC-B; the residual granule porosity increases steadily up to 35% in BHC-A, whereas it goes through a maximum of 35% at 12 weeks and decreases somewhat until 24 weeks in BHC-B. New bone formation within granules appears higher in BHC-A (58% Area) compared to BHC-B (38% area) at 12 weeks. At 24 weeks bone colonization levels off in both cements at about 50% area. Irrespective of the cement matrix composition, beta-TCP granules contribute actively to the conduction of new bone formation.

Augmentation of mechanical properties in osteoporotic vertebral bones--a biomechanical investigation of vertebroplasty efficacy with different bone cements.

Recent clinical trials have reported favorable early results for transpedicular vertebral cement reinforcement of osteoporotic vertebral insufficiencies. There is, however, a lack of basic data on the application, safety and biomechanical efficacy of materials such as polymethyl-methacrylate (PMMA) and calciumphospate (CaP) cements. The present study analyzed 33 vertebral pairs from five human cadaver spines. Thirty-nine vertebrae were osteoporotic (bone mineral density < 0.75 g/cm2), 27 showed nearly normal values. The cranial vertebra of each pair was augmented with either PMMA (Palacos E-Flow) or experimental brushite cement (EBC), with the caudal vertebra as a control. PMMA and EBC were easy to inject, and vertebral fillings of 20-50% were achieved. The maximal possible filling was inversely correlated to the bone mineral density (BMD) values. Cement extrusion into the spinal canal was observed in 12% of cases. All specimens were subjected to axial compression tests in a displacement-controlled mode. From load-displacement curves, the stiffness, S, and the maximal force before failure, Fmax, were determined. Compared with the native control vertebrae, a statistically significant increase in vertebral stiffness and Fmax was observed by the augmentation. With PMMA the stiffness increased by 174% (P = 0.018) and Fmax by 195% (P = 0.001); the corresponding augmentation with EBC was 120% (P = 0.03) and 113% (P = 0.002). The lower the initial BMD, the more pronounced was the augmentation effect. Both PMMA and EBC augmentation reliably and significantly raised the stiffness and maximal tolerable force until failure in osteoporotic vertebral bodies. In non-porotic specimens, no significant increase was achieved.

The effects of calcium phosphate cement particles on osteoblast functions.

Calcium phosphate cements (CPC) are increasingly used in the orthopedic field. This kind of cement has potential applications in bone defect replacements, osteosynthetic screw reinforcements or drug delivery. In vivo studies have demonstrated a good osteointegration of CPC. However, it was also observed that the resorption of CPC could create particles. It is known from orthopedic implant studies that particles can be responsible for the peri-implant osteolysis. Biocompatibility assessment of CPC should then be performed with particles. In this study, we quantified the functions of osteoblasts in the presence of beta-TCP, brushite and cement particles. Two particle sizes were prepared. The first one corresponded to the critical diameter range 1-10 microm and the second one had a diameter larger than 10 microm. We found that CPC particles could adversely affect the osteoblast functions. A decrease in viability, proliferation and production of extracellular matrix was measured. A dose effect was also observed. A ratio of 50 CPC particles per osteoblast could be considered as the maximum number of particles supported by an osteoblast. The smaller particles had stronger negative effects on osteoblast functions than the larger ones. Future CPC development should minimize the generation of particles smaller than 10 microm.

Volume effect on biological properties of a calcium phosphate hydraulic cement: experimental study in sheep.

Injectable calcium phosphate hydraulic cements (CPHC) are a new family of bone substitutes within the class of bone reconstruction biomaterials. In this work, CPHC were tested in two consistencies (preset blocks or liquid paste) in an experimental model of cancellous bone defect in sheep. The defects were eight times larger than those investigated previously in rabbits. Three delays (12, 24, and 52 weeks) were used. Before death, a double label of oxytetracycline and alizarine was made intravenously. The distribution of implants was randomized, histomorphometric evaluation was performed and compared with micrographic observation, and optical microscopy of stained sections was performed either under visible, ultraviolet, or polarized light. The results were compared with spontaneous healing of empty defects and with a control group of normal cancellous bone from sheeps of the same age. No significant difference has been observed between premolded and injected implants. In the sheep model, the degradation and new bone formation rates are three times slower, compared with those observed previously in rabbits. New bone formation increased from 5.9% (12 weeks) up to 11.0% (24 weeks) in the empty defect group. In the cement groups, 28.3% new bone was obtained at 12 weeks, which seemed then to level off (27.8% new bone at 24 weeks). Cement residues appear as radio-opaque cylinders on microradiographs. In all cases, a radiolucent layer was observed at the cement/bone interface at 24 weeks. Stained sections showed the formation of a fibroconnective capsule around the residual cement, which presumably slows down new bone formation. Nevertheless, quantitative bone remodeling was accelerated in the cement group; mineral apposition as well as adjusted apposition rates were higher, and the formation period as well as the mineralization of osteoid tissue were faster compared with empty cavities and controls. These results point to higher osteoblast activity and better exchange with surrounding tissues in the defects filled with cement.

Histological and biomechanical studies of two bone colonizable cements in rabbits.

We have developed two colonizable bone cements: the first is a partially resorbable bisphenol-alpha-glycidyl methacrylate (Bis-GMA)-based cement (PRC) and the second is a calcium phosphate cement (CPC). PRC is composed of aluminous silanized ceramic and particles of a bioresorbable polymer embedded in a matrix of Bis-GMA. CPC consisted of tricalcium phosphate, monocalcium phosphate monohydrate, dicalcium phosphate dihydrate, and xanthane. Both cements were implanted into cavities drilled in rabbit femoral and tibial condyles. After 2, 4, 12, and 24 weeks of implantation, histological observations and biomechanical tests were performed. With CPC, a progressive osteointegration with a concomitant biodegradation in the presence of macrophages were observed. The mechanical study revealed a decrease of the compressive strength until the 4th week, followed by a slight increase. There was a general decrease in the elastic modulus with time. Moreover, by week 4, the histological study showed that the new bone was in direct contact with CPC margins. No inflammation was observed during the observation period. With PRC, the osteointegration as well as the biodegradation were slight, but its compressive strength was higher than that of cancellous bone and CPC (p < 0.05) at all observation periods. Its elastic modulus was greater than that of cancellous bone and CPC until the 4th week, then fell under the values of the cancellous bone.

Raman microspectrometry studies of brushite cement: in vivo evolution in a sheep model.

Calcium phosphate hydraulic cements are promising synthetic bone grafting materials. Brushite-based cements were implanted for 6 and 12 months in the distal condyle of sheep femur, and their in vivo evolution was investigated by Raman microspectrometry. This new technique can probe small volumes in the cubic micrometer range. Its resolution allows a very fine analysis of crystalline changes in calcium phosphate mixtures at the microscopic level. First, Raman spectra of pure brushite, monetite, and beta-tricalcium phosphate (beta-TCP) were recorded, in order to set a data base for the basic components of brushite cements. These spectra show significant differences in the vibration mode v1 for the phosphate ion (988 and 878 cm(-1) for brushite, 988 and 900 cm(-1) for monetite, 968 and 948 cm(-1) for beta-TCP). These differences are strong enough as to allow the qualitative and quantitative analysis of these crystalline phases in the cement. Implanted sheep femur samples were harvested after 24 and 52 weeks post-op, and prepared for Raman analysis in the form of 1-mm-thick sections. Implants at 24 weeks show a core of residual cement isolated from the surrounding bone by fibroconnective tissue. No trace of brushite was detected by micro-Raman analysis in this area, but instead, a mixture of beta-TCP and Type-B carbonated apatite, the latter being very close in composition and structure to the mineral fraction of normal bone in the vicinity of the implant. Implants recovered after 52 weeks show a decrease of the bone/residual cement perimeter, whereas new trabeculations are formed in the implanted zone; the small amounts of residual cement still present are substantially transformed into Type-B carbonated apatite containing small amounts of proteins. In the same area, some beta-TCP particles are also detected showing that, contrary to brushite, the excess beta-TCP originally present in the cement is not completely metabolized. In the implanted zone already converted into trabecular bone, Raman microspectrometry shows the characteristic spectrum of normal bone.

Reinforcement of osteosynthesis screws with brushite cement.

The fixation of osteosynthesis screws remains a severe problem for fracture repair among osteoporotic patients. Polymethyl-methacrylate (PMMA) is routinely used to improve screw fixation, but this material has well-known drawbacks such as monomer toxicity, exothermic polymerization, and nonresorbability. Calcium phosphate cements have been developed for several years. Among these new bone substitution materials, brushite cements have the advantage of being injectable and resorbable. The aim of this study is to assess the reinforcement of osteosynthesis screws with brushite cement. Polyurethane foams, whose density is close to that of cancellous bone, were used as bone model. A hole was tapped in a foam sample, then brushite cement was injected. Trabecular osteosynthesis screws were inserted. After 24 h of aging in water, the stripping force was measured by a pull-out test. Screws (4.0 and 6.5 mm diameter) and two foam densities (0.14 and 0.28 g/cm3) were compared. Cements with varying solid/liquid ratios and xanthan contents were used in order to obtain the best screw reinforcement. During the pull-out test, the stripping force first increases to a maximum, then drops to a steady-state value until complete screw extraction. Both maximum force and plateau value increase drastically in the presence of cement. The highest stripping force is observed for 6.5-mm screws reinforced with cement in low-density foams. In this case, the stripping force is multiplied by 3.3 in the presence of cement. In a second experiment, cements with solid/liquid ratio ranging from 2.0 to 3.5 g/mL were used with 6.5-mm diameter screws. In some compositions, xanthan was added to improve injectability. The best results were obtained with 2.5 g/mL cement containing xanthan and with 3.0 g/mL cements without xanthan. A 0.9-kN maximal stripping force was observed with nonreinforced screws, while 1.9 kN was reached with reinforced screws. These first results are very promising regarding screw reinforcement with brushite cement. However, the polyurethane foam model presents noninterconnected porosity and physiological liquid was not modelized.

Gentamicin-loaded hydraulic calcium phosphate bone cement as antibiotic delivery system.

A hydraulic calcium phosphate cement made of beta-tricalcium phosphate [beta-Ca3(PO4)2], monocalcium phosphate monohydrate [Ca(H2PO4)2-H2O], and water was used as a delivery system for the antibiotic gentamicin sulfate (GS). GS, added as powder or as aqueous solution, was very beneficial to the physicochemical properties of the cement. The setting time increased from 2 to 4.5 min with 3% (w/w) GS and then slowly decreased to 3.75 min with 16% (w/w) GS. The tensile strength increased from 0.4 to 1.6 MPa with 16% (w/w) GS. These effects were attributed to the presence of sulfate ions in GS. The release of GS from the cement was measured in a pH 7.4 phosphate-buffered saline solution at 37 degrees C by USP paddle method. Factors such as cement porosity, GS content and presence of sulfate ions or polymeric additives were investigated. The amount of GS released was roughly proportional to the square root of time up to approximately 50% release. Afterwards, the release rate markedly slowed down to zero. In all but two cement formulations, the total dose of GS was released within 7 days, indicating that no irreversible binding occurred between the cement paste and the antibiotic. When small amounts of hydroxypropylcellulose or poly(acrylic acid) were added to the cement, the maximum fraction released was a few percent lower than the total GS dose, suggesting some binding between the polymer and GS. The GS release rate was strongly influenced by the presence of sulfate ions in the cement paste and by the cement porosity. The higher the sulfate ion content of the cement paste, the lowe the GS release rate. This influence was attributed to the finer cement micro-structure induced by the presence of sulfate ions. Furthermore, when the initial cement porosity was increased from 38 to 69%, the release rate almost tripled (0.16 to 0.45 h-1/2). Finally, the biological activity of GS in the cement was maintained, as measured by assaying the release medium.

Composition effects on the pH of a hydraulic calcium phosphate cement.

The pH of a hydraulic calcium phosphate cement (HCPC) made of monocalcium phosphate monohydrate (Ca(H2PO4)2.H2O; MCPM), beta-tricalcium phosphate (beta-(Ca3(PO4)2; beta-TCP) and water was measured as a function of reaction time and composition at room temperature. During setting, the cement pH varies from very acidic pH values, i.e., 2.5, to almost neutral pH values, i.e., 6. The cement pH profile significantly depends on the initial cement composition. However, all profiles are characterized by a sharp initial decrease of the pH due to the dissolution of MCPM crystals and the precipitation of dicalcium phosphate dihydrate (CaHPO4. 2H2O; DCPD) crystals. With an excess of MCPM, the final pH stays low, and its value can be predicted from the initial composition of the cement and solubility data. With an excess of beta-TCP, the end pH is close to 5, which is much lower than 5.9, the value predicted by calculation. Results suggest that the difference may be due to the presence of impurities in the cement. Replacing MCPM by phosphoric acid renders the cement paste very acidic for the initial 30 s, but then the pH profile follows that obtained with MCPM. Adding pyrophosphate ions into the cement paste postpones the position of the pH minimum. The delay, which is proportional to the concentration of pyrophosphate ions, is thought to be due to the inhibiting action of pyrophosphate ions on the precipitation of DCPD crystals.

Synthesis of bioactive coatings on Ti substrates using glass enamel.

Bioactive coatings on titanium can be prepared by dispersing hydroxyapatite particles into a borosilicate glass enamel doped with TiO2. Adhesion of the coating is obtained when the titanium substrate has been preoxidized before enameling in such a way as to form a continuous TiO2-layer. Adhesion results from the diffusion of this TiO2-layer into the liquid glass during enameling. The evolution of the diffusion zone was followed by scanning electron microscopy and electron probe microanalysis. Adhesion disappears after a critical time of firing which corresponds to the completion of the dissolution of the TiO2-layer by the glass. The borosilicate glass wets the hydroxyapatite particles and adheres strongly to them after cooling.

Chemical analysis of renal stones from 377 Belgian patients by using qualitative or quantitative methods.

The renal stones of 377 patients from the Brussels' area have been studied by chemical methods; 239 stones were submitted to qualitative analysis and the other 138 to a quantitative analysis. The results of the study demonstrate that, in Belgium as well as in other Western countries like the USA and Great Britain, calcium oxalate, calcium phosphate and magnesium ammonium phosphate are, in decreasing order of frequency, the major constitutents of renal calculi. The fact that calcium oxalate-containing stones are mainly found in men associated with sterile urine, and magnesium ammonium phosphate stones in women associated with urinary infection, is confirmed in the present series. The percentage of uric acid-containing stones is similar to that in the USA and Great Britain, but lower than that observed in several European countries including France, Spain, Germany, Czechoslovakia and Sweden. A small group of Mediterranean patients living in Belgium show no specific pattern, suggesting that the formation of calculi could be more dependent upon environmental than upon ethnic factors.