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Bonelike apatite formation on ethylene-vinyl alcohol copolymer modified with silane coupling agent and calcium silicate solutions.

An ethylene-vinyl alcohol copolymer (EVOH) was treated with a silane coupling agent and calcium silicate solutions, and then soaked in a simulated body fluid (SBF) with ion concentrations approximately equal to those of human blood plasma. A smooth and uniform bonelike apatite layer was successfully formed on both the EVOH plate and the EVOH-knitted fibers in SBF within 2 days. Part of the structure of the resulting apatite-EVOH fiber composite was similar to that of natural bone. If this kind of composite can be fabricated into a three-dimensional structure similar to natural bone, the resultant composite is expected to exhibit both mechanical properties analogous to those of natural bone and bone-bonding ability. Hence, it has great potential as a bone substitute.

Apatites↗

In vivo absorption of porous apatite- and wollastonite-containing glass-ceramic.

The behavior of porous apatite- and wollastonite-containing glass-ceramic (AW) in the bone marrow cavity was investigated. Cylinders of porous AW (4 mm in diameter and 20 mm long, mean porosity of 70% and mean pore diameter of 200 microm) were implanted into the bone marrow cavity of rabbit femurs, and analyzed by chronological radiograms and by scanning electron microscopy one, three, six, and 12 months later. The pores of porous AW are interconnected and homogeneously distributed, and its compressive strength is nearly equal to that of human cancellous bone. Bone formed in the pores at the center of the material by one month and bonded to the material directly. The volume of newly formed bone in the material pores reached a peak at three months, and decreased gradually after six months. The trabecular structures of AW were gradually remodeled by newly formed bone, while AW-bone bonding was maintained during bone remodeling and material absorption. AW was absorbed continuously, and at six and 12 months the residual material corresponded to about 64 and 30% of the starting material, respectively. Porous AW may therefore be useful as an absorbable bone substitute.

Absorbable Implants↗

Autogenous injectable bone for regeneration with mesenchymal stem cells and platelet-rich plasma: tissue-engineered bone regeneration.

We have attempted to regenerate bone in a significant osseous defect with minimal invasiveness and good plasticity, and to provide a clinical alternative to autogenous bone grafts. Platelet-rich plasma (PRP) may enhance the formation of new bone and is nontoxic, nonimmunoreactive, and accelerates existing wound-healing pathways. We have used a combination of PRP as an autologous scaffold with in vitro-expanded mesenchymal stem cells (MSCs) to increase osteogenesis, compared with using the scaffold alone or autogenous particulate cancellous bone and marrow (PCBM). The newly formed bones were evaluated by radiography, histology, and histomorphometric analysis in the defects at 2, 4, and 8 weeks. According to the histological observations, the dog MSCs (dMSCs)/PRP group had well-formed mature bone and neovascularization compared with the control (defect only), PRP, and PCBM groups at 2 and 4 weeks. Histometrically, at 8 weeks newly formed bone areas were 18.3 +/- 4.84% (control), 29.2 +/- 5.47% (PRP), 61.4 +/- 3.38% (PCBM), and 67.3 +/- 2.06% (dMSCs/PRP). There were significant differences between the PCBM, dMSCs/PRP, and control groups. These results demonstrate that the dMSCs/PRP mixture is useful as a osteogenic bone substitute.

Animals↗

Durability of oscillating bone saw blades.

The purpose of this study was to measure the durability of disposable saw blades using a standard bone-cutting experimental model that can be replicated in any laboratory. Using a constant cutting force, the bone-cutting efficiency was determined by recording the maximum cutting temperature and the rate of material removal. In this experimental study, porcine femurs as well as a synthetic bone substitute were used as the test samples for measuring cutting efficiency. Repeated cutting of these test materials did not significantly alter the performance of the disposable bone-cutting blades.

Animals↗

Biomimetic porous scaffolds with high elasticity made from mineralized collagen--an animal study.

Histological investigations of a new hydroxyapatite-collagen composite material were carried out to evaluate its possible suitability as a bone substitute. The three-dimensional scaffolds made from biomimetically mineralized collagen exhibit an interconnecting pore structure and elastic mechanical properties. They were implanted into the subcutaneous tissue and bone defects made in the femur of rats and harvested with the surrounding tissue at 1, 2, 4, 8, and 12 weeks after surgery. The materials implanted in the subcutaneous tissue were covered by fibrous connective tissue with a slight inflammatory response, and many foreign-body giant cells were observed on the surface of the scaffolds. Most of the material implanted in the subcutaneous tissue was resorbed at 8 weeks by phagocytosis. In the bone defects, new bone formation was observed on the surface of the material at 1 week. New bone increased with time, and osteoclasts were seen on the surface of the scaffolds at 2 weeks. Resorption and replacement by new bone of many parts of the materials implanted in the femur were observed by 12 weeks. These responses occurred faster than those of other hydroxyapatite-collagen composites. The results suggested that the new biomimetically mineralized collagen scaffolds were suitable as an implant material for bone-tissue reconstruction.

Animals↗

Bonding strength of bonelike apatite layer to Ti metal substrate.

Our previous study showed that titanium metal forms a bonelike apatite layer on its surface in simulated body fluid when it was subjected to NaOH and heat treatments to form a sodium titanate hydrogel or amorphous sodium titanate surface layer. In the present study, bonding strength of the apatite layer formed on the titanium metals to the substrates were examined under tensile stress, in comparison with those of the apatite layers formed on Bioglass 45S5-type glass, dense sintered hydroxyapatite, and glass-ceramic A-W, which are already clinically used. The NaOH-treated titanium metals showed higher bonding strength of the apatite layer to the substrates, which was maximized by heat treatments at 500 and 600 degrees C, than all the examined bioactive ceramics. It is believed that bioactive metals thus obtained are useful as bone substitutes, even under load-bearing conditions.

Apatites↗

Coral as a carrier for recombinant human bone morphogenetic protein-2.

By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage growth was induced in the pores or on the surface of the implants at one week, woven bone at three week and lamellar bone with bone marrow at six week, and coral was absorbed partially. The induced formation of endochondral bone was time-related and rhBMP-2 dose-related. The results of this study indicate that the composite possesses a superior ability of osteogenesis, and coral acts as one of the most suitable rhBMP-2 slowrelease carriers currently available. The composite will be a new type of bone substitute to be used in orthopaedics and maxillofacial surgery.

Animals↗

Development of a beta-type Ti-12Mo-5Ta alloy for biomedical applications: cytocompatibility and metallurgical aspects.

Ti-based biocompatible alloys are especially used for replacing failed hard tissue. Some of the most actively investigated materials for medical implants are the beta-Ti alloys, as they have a low elastic modulus (to inhibit bone resorption). They are alloyed with elements such as Nb, Ta, Zr, Mo, and Fe. We have prepared a new beta-Ti alloy that combines Ti with the non-toxic elements Ta and Mo using a vacuum arc-melting furnace and then annealed at 950 degrees C for one hour. The alloy was finally quenched in water at room temperature. The Ti-12Mo-5Ta alloy was characterised by X-ray diffraction, optical microscopy, SEM and EDS and found to have a body-centred-cubic structure (beta-type). It had a lower Young's modulus (about 74 GPa) than the classical alpha/beta Ti-6Al-4V alloy (120 GPa), while its Vickers hardness remained very high (about 303 HV). This makes it a good compromise for a use as a bone substitute. The cytocompatibility of samples of Ti-12Mo-5Ta and Ti-6Al-4V titanium alloys with various surface roughnesses was assessed in vitro using organotypic cultures of bone tissue and quantitative analyses of cell migration, proliferation and adhesion. Mechanically polished surfaces were prepared to produce unorientated residual polished grooves and cells grew to a particularly high density on the smoother Ti-12Mo-5Ta surface tested.

Animals↗

Synthesis of bioactive PMMA bone cement via modification with methacryloxypropyltri-methoxysilane and calcium acetate.

Bone cement consisting of polymethylmethacrylate (PMMA) powder and methylmethacrylate (MMA) liquid is clinically used for fixation of implants such as artificial hip joints. However, it does not show bone-bonding ability, i.e., bioactivity. The lack of bioactivity would be one of factors which cause loosening between the cement and the implant. The present authors recently showed the potential of bioactive PMMA-based bone cement through modification with gamma-methacryloxypropyltrimethoxysilane (MPS) and calcium acetate. In this study, the effects of the kinds of PMMA powder on setting time, apatite formation and compressive strength were investigated in a simulated body fluid (Kokubo solution). The cement modified with calcium acetate calcined at 220 degrees C could set within 15 min when the PMMA powder had an average molecular weight of 100,000 or less. The addition of calcium acetate calcined at 120 degrees C in the PMMA powder required a much longer period for setting. The modified cements formed an apatite layer after soaking in the Kokubo solution within 1 day for cement starting from PMMA powder with a molecular weight of 100,000 or less. Compressive strengths of the modified cements were more than 70 MPa for cements starting from 100,000 and 56,000 in molecular weight. After soaking in Kokubo solution for 7 days, the modified cement consisting of PMMA powder of 100,000 in molecular weight showed a smaller decrease in compressive strength than that consisting of 56,000 in molecular weight. These results indicate that bioactive PMMA cement can be produced with appropriate setting time and mechanical strength when PMMA powders with a suitable molecular weight are used. Such a type of design of bioactive PMMA bone cement leads to a novel development of bioactive material for bone substitutes.

Acetates↗

Sinus floor augmentation using a composite graft of bone morphogenic protein-2 and allogenic cancellous bone (Puros): case report.

Reconstruction of the atrophic maxilla is a difficult task. The gold standard for such reconstruction is autogenous bone. Presently, many excellent products are available to the dental surgeon to facilitate alveolar reconstruction in the absence of autogenous bone. This study describes the use of bone morphogenic protein in combination with allogenic bone substitute (Puros) to reconstruct the maxilla in preparation for dental implant placement.

Alveolar Bone Loss↗

Localisation of osteogenic and osteoclastic cells in porous beta-tricalcium phosphate particles used for human maxillary sinus floor elevation.

We and others have shown earlier that porous beta-tricalcium phosphate (TCP) (Cerasorb) can be used in patients to augment the maxillary sinus floor prior to placement of oral dental implants. To better understand the transformation of TCP particles into bone tissue, we analyse here the appearance of cells with osteogenic or osteoclastic potential in relation to these particles. In biopsies taken at 6 months after sinus floor augmentation we observed bone growth into the TCP particles but also replacement by soft connective tissue. To identify possible osteoprogenitor cells in this tissue, histological sections were immunostained with an antibody to Runx2/Cbfa1, an essential and early transcription factor for osteoblast differentiation. The osteogenic potential of cells was further confirmed by immunostaining for bone sialoprotein (BSP) and osteopontin (OPN). Other sections were stained for Tartrate Resistant Acid Phosphatase (TRAP) activity to identify cells with osteoclastic capacity. Runx2/Cbfa1 positive connective tissue cells were found in abundance throughout and around the TCP particles, even at a distance of several millimetres from the maxillary bone surface. About 95% of the cells found within TCP particles stained positive for Runx2/Cbfa1. Fewer cells stained positive for BSP and OPN, suggesting more mature osteoblastic properties. Mono- and binucleate TRAP-positive cells, but no multinucleate TRAP-positive osteoclasts, were found in the soft tissue infiltrating the TCP and at the surface of the TCP particles. Both the Runx2/Cbfa1 positive and the TRAP-positive cells decreased apically with increasing vertical distance from the maxillary bone surface. This data suggests that the TCP particles attract osteoprogenitor cells that migrate into the interconnecting micropores of the bone substitute material by 6 months. The lack of large multinucleate TRAP positive cells suggests that resorption of the TCP material by osteoclasts plays only a minor role in its replacement by bone. Chemical dissolution, possibly favoured by a high cell metabolism in the particles, seems the predominant cause of TCP degradation. The abundance of Runx2/Cbfa1 positive cells would indicate that with a greater time of healing there will be further bone deposition into these particles.

Bone Substitutes↗

Influences of vascularization and osteogenic cells on heterotopic bone formation within a madreporic ceramic in rats.

Research in biomaterials for bone reconstruction has led to elaborate osteogenic composites that combine porous ceramics with bone marrow stromal cells. The aim of this study was to evaluate the influence of direct vascularization of such composites on osteogenesis and the ability to produce a vascularized bone substitute transplant in an ectopic muscular site. Sixty-four coralline biomaterials were implanted in 32 Fisher rats under four conditions: (1) alone (reference group M, n = 16), (2) coated with bone marrow stromal cells (group MC, n = 16), (3) combined with a vascular pedicle (group MV, n = 16), or (4) coated with bone marrow stromal cells and combined with a vascular pedicle (MCV group, n = 16). The number of vessels in the pores (vessel-pore ratio) of the implants and the proportion of pores showing bone ingrowth (bone-pore ratio) were measured at 2, 4, 6, and 8 weeks on four implants of each group. Compared with the reference group, angiogenesis was higher when the biomaterial was combined with a vascular pedicle or was coated with osteoprogenitor cells. The association of both vascular pedicle and osteoprogenitor cells increased vascularization by 60 percent (p = 0.003) and osteogenesis by 62 percent (p < 0.001). A combination of both vascular pedicle and bone marrow osteoprogenitor cells in coralline implants enhances neovascularization and osteogenesis after implantation in ectopic intramuscular sites to a greater extent than either does alone.

Animals↗

Injectable bone using chitosan-alginate gel/mesenchymal stem cells/BMP-2 composites.

AIM: Several injectable materials have been used as osteogenic bone substitutes. However, none has gained universal acceptance. This study was performed to investigate whether or not chitosan-alginate gel/mesenchymal stem cells/bone morphogenetic protein-2 composites are potentially injectable materials for new bone formation. MATERIAL AND METHODS: The composites were injected into the subcutaneous space on the dorsum of nude mice to investigate new bone tissue formation. The composites were examined histologically over a 12-week period. RESULTS: The composites injected into the mouse were able to stimulate new bone formation, which was trabecular in type. CONCLUSIONS: This study showed that chitosan-alginate gel/MSCs/BMP-2 composites could become clinically useful injectable materials to generate new bone.

Alginates↗

The use of nanocrystalline hydroxyapatite for the reconstruction of calcaneal fractures: Preliminary results.

The purpose of this investigation was to evaluate the feasibility of a nanocrystalline hydroxyapatite compound in the treatment of calcaneal fractures with osseous defects after reduction. The study included 21 patients, representing 24 closed intraarticular calcaneus fractures with large defects remaining after operative reduction. All cases were supplemented with the hydroxyapatite bone substitute and stabilized with a calcaneal honeycomb plate. Radiographs were taken at 6 weeks, 12 weeks, 6 months, and 1 year postoperatively, with specific attention given to measurement of Gissane's angle, Böhler's angle, and calcaneal height. A postoperative subjective and objective evaluation of the fracture, using the Creighton Nebraska Health Foundation scale, was performed 1 year postoperatively. Böhler's angle improved from a mean 8.6 +/- 5.3 degrees preoperatively to an immediate postoperative mean result of 31.5 +/- 6.5 degrees and 27.7 +/- 8.6 1 year postoperatively. The mean Creighton-Nebraska functional score was 86 +/- 10 at the 1-year follow-up evaluation. These results suggest that open reduction with plate fixation combined with nanocrystalline hydroxyapatite augmentation presents a good and reliable surgical technique for treatment of calcaneus fractures.

Adult↗

[Filling of bone defects using biphasic macroporous calcium phosphate ceramic. Apropos of 23 cases].

PURPOSE OF THE STUDY: The authors report their experience with the use of a biphasic macroporous calcium phosphate bone substitute. MATERIALS AND METHODS: In 23 cases (22 patients) a biphasic macroporous calcium phosphate ceramic was used to fill a pathological bone defect. The ceramic used in this study was a macroporous (400 to 600 mu) component consisting of 60 per cent hydroxyapatite and 40 per cent beta-tricalcium phosphate. It was in the form of granules (2 to 3 mm), sticks (20 x 5 x 5 or 10 x 5 x 5 mm) or custom made blocks. In 6 cases, the ceramic was used alone; in 12 cases with autologous bone marrow and in 5 cases with autologous cancellous bone grafts. In 14 cases, the bone defect was due to conservative treatment of a benign tumor, in 3 cases due to aseptic post-traumatic non union, in 3 cases due to wide resection for malignant tumors of the pelvis and in 3 cases following osteotomy. Post operative assessment was made from clinical, radiographic and histological findings. RESULTS: 2 patients died 6 and 8 months post operatively and 2 were lost to follow up at 2 and 5 months with both having good clinical and radiographic results when last seen. For the remaining 19 cases, the average follow-up was 20 months (from 6 to 62 months). No local, regional or general deleterious effects were noted. Radiologically the bone ceramic junction healed in all cases except 2 within 3 months. In these last two cases, healing required 6 and 7 months. No radiolucent line appeared around the ceramic. No stress fractures occurred in the substitute. Histologically, 3 biopsies showed new bone formation throughout the ceramic with apposition of a well differentiated lamellar bone directly apposed to the ceramic. DISCUSSION: Animal experimentations have proven the interest of similar ceramics: macroporosity enhances bone rehabilitation and the biphasic characteristics associate the advantages of slow resorption of hydroxyapatite and more easily resorbed beta-calcium phosphate. No deleterious clinical, radiographical or histological effects were observed, confirming the biocompatibility of this substitute. Despite the poor mechanical properties of this macroporous ceramic before implantation, good clinical and radiographic results suggest improvement of these properties in the composite new formed bone-ceramic after implantation. CONCLUSION: We believe that macroporous biphasic ceramic is a good substitute for use in bone defects when good primary mechanical stability and contact with the host bone are present. Further clinical and experimental studies are necessary to determine the limits of such a substitute in terms of volume and to control its mechanical properties following implantation.

Adolescent↗

Tissue engineering and skeletal diseases.

Tissue engineering, a cross between the science of the living organism and that of engineering, aims to replace, maintain or improve human tissue functions, by means of tissue substitutes containing living elements. Thus, it is about production of artificial tissue, using (alone or in combination) cells, matrix or bioactive factors. Their association gives rise to a hybrid biomaterial combining biological components (cells, growth factors or adhesion proteins) and materials (polymers, ceramics). The applications are wide-ranging, from the skin, to the liver, or to the cornea as well as to the locomotor system. Bone tissue engineering has advanced the most in this field, partly because of the progress made by research into bone substitutes, although cartilage and tendons are also concerned. This technology requires cell culture (committed cells or more often bone marrow stem cells), biomaterials (porous materials with controlled architecture and cements), growth factors (such as 'Bone Morphogenetic Proteins'), the proteins implicated in cell adhesion (such as fibronectin or the aminoacid sequences specifically recognised by integrin subunits) or gene therapy (notably using transfected stem cells). Tissue engineering and regenerative stimulation of tissue are now booming on experimental and industrial levels and clinical applications are increasingly numerous. Considering the potential of these technologies, they should continue to develop widely.

Biomedical Engineering↗

In vitro attachment of osteoblast-like cells to osteoceramic materials.

OBJECTIVE: The objective of this work was to examine osteoblast-like cell attachment and morphology in vitro to osteoceramic materials with three different surface morphologies. METHODS: Osteoceramic composite disks were fabricated from tricalcium phosphate and magnesium-aluminate spinel (MgAl2O4) in a 50 vol% ratio. The disks were prepared with three different surface morphologies, including as-fired (irregular), etched (rough), or polished through 1 mm diamond paste (smooth). Osteoblast-like cell cultures were plated onto the prepared disks for 2 h, and the number of attached cells was determined. ANOVA and Student Newman-Kuels tests were used to test for significant differences in cell attachment (p < 0.05). SEM was used to visually evaluate the nature of the cellular adaptation on the osteoceramic surfaces. RESULTS: Some additional surface roughening resulted from the interaction between the osteoceramic disks and the biological culture media during the attachment assay. A statistically larger number of cells was found to be attached to the etched osteoceramic surfaces compared to the as-fired and polished osteoceramic surfaces or the tissue culture plastic control. Cellular adaptation was extensive on all three osteoceramic surfaces at 2 h. SIGNIFICANCE: These results are consistent with previous in vivo work and continue to support the hypothesis that osteoceramic materials have potential for implants and bone substitute materials.

Aluminum Compounds↗

[Experimental study on the reconstruction of mandibular defects with a new bioactive artificial bone nano-hydroxyapatite/polyamide-66 in dogs].

OBJECTIVE: This study was designed to investigate reconstruction of segmental defect in the mandible using a new bionic materials of nano-hydroxyapatite -polyainides-66 (n-HA/PA66). METHODS: Two defects (15 mm x 10 mm x 5 mm) were created in the mandibular bodies of dogs. One of defects was reconstructed with n-HA/PA66, another not repaired as a blank control. At 2, 4, 8, 12, 16 weeks after operation. Evaluation of effects of n-HA/PA66 on reconstruction of the mandibular defects was carried out by means of radiography and histology. RESULTS: From 2 to 8 weeks after operation, some fiber tissue grew into the space between n-HA/PA66 and mandibular bone. The ossification was observed at 12 weeks post-operation. At 16 weeks, the n-HA/PA66 was connected directly to the mandibular bone by the newborn bone. CONCLUSIONS: The new artificial bone of n-HA/PA66 has the effects of osteoconduction and osteoinduction, with a good biocompatibility and is an ideal bone substitute material for reconstruction of mandibular defect.

Animals↗