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Tissue reactions to particles of bone-substitute materials in intraosseous and heterotopic sites in rats: discrimination of osteoinduction, osteocompatibility, and inflammation.

Two rat models were used to characterize tissue-specific reactions to particles of bone-substitute materials: one for osteocompatibility in a healing tibial wound and the other in a heterotopic, subcutaneous site. Small, unicortical tibial wounds in rats healed spontaneously, beginning with the rapid proliferation of intramedullary woven bone. That temporary bone was resorbed by osteoclasts and finally, the cortical wound was healed with lamellar bone and the medullary space was repopulated with marrow. When various particulate materials were implanted into fresh wounds, three types of reactions were observed. (1) Demineralized bone powder (DBP) and non-resorbable calcium phosphate (nrCP) were incorporated into the reactive medullary and cortical bone. (2) Polymethylmetlhacrylate (PMMA) particles were surrounded with a fibrous layer, but did not impair bone healing. (3) Polyethylene (PE) shards and resorbable calcium phosphates (rCPs) were inflammatory and inhibited osseous repair. Subcutaneous sites showed osteoinductive, fibrotic, or inflammatory responses to these materials. Only DBP induced endochondral osteogenesis subcutaneously. The nrCP evoked a fibrous reaction. In contrast, rCPs, PMMA, and PE shards generated inflammatory reactions with each particle being surrounded by fibrous tissue and large multinucleated giant cells. In conclusion, only DBP showed osteoinductive as well as osteocompatible properties. The nrCP was osteocompatible. The rCPs stimulated various degrees of inflammatory responses. PMMA was osteocompatible and did not interfere with the bone healing process. PE was not osteocompatible and generated foreign body reactions in both sites. Use of the two sites distinguishes osteoinductive, osteocompatible, and inflammatory properties of particles of bone-substitute materials.

Absorbable Implants↗

[Development of a model which makes it possible to test injectable bone substitutes and evaluation of a calcium phosphate cement].

New percutaneous filling techniques are beginning to be used in bone tumor pathology. The purpose of this study was to develop a reproducible model for testing new injectable bone substitutes. A closed cancellous bone defect was created in the distal femoral condyles of rabbit. Bone defect was filled by a percutaneous injection. Several situations were tested: spontaneous evolution of unfilled cavities, evolution of cavities filled either with a conventional orthopedic cement (PMMA), either with a mixed collagen-hydroxyapatite material, either with a biomaterial derived from the vegetal protein zein, either with a calcium phosphate cement brushite type. The new bone formation was quantified in the defect for each group in which several delays were tested. A bone defect of reproducible size was obtained consistently. A partial bone formation was observed in the unfilled group. The best bone formation rate was obtained in the phosphate cement group. This model showed the interest of injectable biomaterials and more precisely the interest of the phosphate calcium cements as brushite.

Animals↗

Resorbability of bone substitute biomaterials by human osteoclasts.

Third generation biomaterials are being designed with the aim that once implanted they will help the body to heal itself. One desirable characteristic of these materials in bone is their ability to be remodeled, i.e. that osteoclasts resorb the material and it is subsequently replaced by newly formed bone through osteoblastic activity. So far the only way to test this biological property of bone substitutes are animal experiments with all their limitations like ethics, costs and limited transferability to man. The present study was designed, to develop a human in vitro assay, allowing to generate human osteoclasts directly on the biomaterial. The assay was validated using calcium phosphate cement and PMMA as biomaterials. Quantification was performed by raster electron microscopy and computer assisted image analysis. Dentin was used as internal standard. Our assay shows iso-bone resorbability of calcium phosphate cement in comparison to unresorbable PMMA cement. Both current clinical orthopedic practice and future skeletal engineering may profit from the availability and use of a test system for the assessment of resorption quality. The assay presented here allows to address this question of resorbability and to select the best materials for the use as bone substitutes in specific patients.

Absorbable Implants↗

Comparative study of beta-tricalcium phosphate mixed with platelet-rich plasma versus beta-tricalcium phosphate, a bone substitute material in dentistry.

Animal experiments were carried out with osteoconductive bone substitute beta-tricalcium phosphate (beta-TCP), with the aim of assessing the effects of the growth factors synthesised by thrombocytes on the speed of beta-TCP incorporation and on the quality of newly formed bone. The question to be answered was the extent to which platelet-rich plasma (PRP) accelerated the resorption of beta-TCP and the formation of new bone. Two teeth were removed symmetrically from each side of the mandible of 12 Beagle dogs; the resulting cavities were filled on one side with beta-TCP alone, and on the other side with a mixture of beta-TCP + PRP (obtained from autologous blood). The quality of the newly formed bone and the effects of this PRP were studied by histological and histomorphometric methods. In week 6, bone formation was already more effective when PRP was applied in comparison with beta-TCP alone, and in week 12 the growth was significantly greater. The results demonstrate that the use of PRP accelerates the remodelling of new bone created by beta-TCP.

Animals↗

[Bone substitute with growth factor. Preliminary clinical cases for cranio- and maxillo-facial indications].

Several biological materials have been analyzed in combination with osteo-inductive growths factors to determine whether such a system can replace bone grafting in surgical practice. Efforts have been aimed at the discovery of the best carriers and delivery systems. We present the results of the surgical treatment of 11 cranio-maxillo-facial defects in 9 patients using a combination of natural coral skeleton (NCS in blocks or granules), human fibrin glue and transforming growth factor beta-1 (TGF-beta 1) as a composite bone substitute. Three patients were initially excluded because of early extrusion of the materials due to a technical error. Clinical and radiological evaluation was performed in all cases, with the patient acting as his own control. Clinical firmness and radiological mineralization occurred in three quarters of cases. New bone formation was confirmed histologically in two of these patients. Clinically the initial results remained stable over a three years follow-up with staged surgical procedures performed on a number of patients. None of the patients suffered any detrimental effect from implantation of the bone substitute. Although the numbers in these series are limited, the association of TGF-beta 1, human fibrin glue and NCS represented an interesting step, although the clinical results could be improved. Important factors in the success of this technique appeared to be stabilisation of the biological materials, quality and asepsis of the surrounding tissue and the dose of growth factor.

Acrocephalosyndactylia↗

[Non-invasive evaluation of an injectable bone substitute].

Despite the increasing number of techniques for the preservation of bone ridges after dental avulsion, no precise evaluation of alveolar filling has been performed to date. The criteria of available measurement techniques (probes, retroalveolar or panoramic radiography, and lateral teleradiography) are not sufficiently reliable and precise. This study investigated the reliability of evaluation based on CT images in comparison with retroalveolar radiography (the most precise radiographic technique, providing standardised images), direct measurements, and images obtained in scanning electron microscopy. After a preliminary investigation ex vivo, a study was performed in vivo on three beagles. Mandibular premolars were extracted, and the corresponding alveoli were filled with an injectable bone substitute composed of a calcium phosphate mineral load associated with hydroxypropyl methylcellulose. Measurements performed on CT images relative to visual and automatic detection of density changes and studies of density curves provided better precision than those obtained by retroalveolar radiography.

Animals↗

Comparing the osteogenic capacities of bone substitutes: hydroxyapatite, high-density porous polyethylene, and bone collagen: a biochemical and histological analysis.

Several inorganic materials have been shown previously to hold some osteogenic capacity. The purpose of this study is to compare the bone-forming abilities of hydroxyapatite ceramic, high-density porous polyethylene, and bone collagen within the periosteal island flap of rabbit tibia using histological and biochemical analysis. With this goal, four discrete experimental groups were formed, each comprising 22 New Zealand male rabbits. A sac was created on each rabbit tibial periosteum flap in each of the groups, and each of the previously mentioned materials was placed within this sac separately. One of these groups was thought as a control group without any material being placed inside the periosteal sac. Biopsies were taken at weeks 1, 2, 4, and 8 for biochemical analysis and at weeks 2 and 8 for histological evaluation. Neo-osteogenesis was evaluated quantitatively by determination of alkaline phosphatase and osteocalcin levels biochemically as well as by the percentage of new bone formation inside the periosteal sac histologically. Results show statistically that the osteogenic effect of high-density porous polyethylene is greater than that of the other materials used in this study (P < 0.05).

Analysis of Variance↗

Injectable bone substitute using a hydrophilic polymer.

We studied a new injectable biomaterial for bone and dental surgery consisting of a hydrophilic polymer as matrix and bioactive calcium phosphate (CaP) ceramics as fillers. This material is composed of complex fluids whose flow is determined by the laws of rheology. We investigated the macromolecular effects on this composite in a tube. The stability of the polymer and the mixture is essential to the production of a ready-to-use injectable biomaterial. These flow properties are necessary to obtain CaP bioactivity in a dental canal or bone defect during percutaneous surgery. Macromolecules provide spaces between CaP ceramic granules and facilitate the role of the biological agents of bone substitution.

Animals↗

Physical and mechanical properties evaluation of Acropora palmata coralline species for bone substitution applications.

The search for ideal materials for bone substitution has been a challenge for many decades. Numerous natural and synthetic materials have been studied. For this application, exoskeletons of coral have been considered a good alternative given its tendency to resorption, biocompatibility and similarity to the mineral bone phase. Very few studies of these materials consider a detailed analysis of the structure-property relationship. The purpose of this work was to carry out the microstructural characterization of a coralline species named Acropora palmata and the determination of the mechanical and physico-chemical properties. Measurements of hardness, compressive strength, bulk density and apparent porosity were performed. From these results it was determined that this marine coral species could be an alternative xenograft due to its mechanical properties and osteoconductive nature.

Journal Article↗

Composites made of rapidly resorbable ceramics and poly(lactide) show adequate mechanical properties for use as bone substitute materials.

Porous composite materials made of poly(L, DL-lactide) and a ceramic component, alpha-tricalcium phosphate (alpha-TCP) or one of the rapidly resorbable glass ceramics, GB9N or GB14N, respectively, were developed to be used as bone substitutes. The present article describes the mechanical properties and the in vitro degradation characteristic of the different composite materials. The yield strength, the elastic modulus, and the molecular weight were measured after in vitro degradation up to 78 weeks. The initial strengths of the alpha-TCP composite (12.5 +/- 0.7 MPa) was higher than that of the GB9N and GB14N composites (8.3 +/- 0.2 MPa and 10.9 +/- 0.2 MPa, respectively). The initial elastic moduli of the composites were between 450 and 650 MPa. The mechanical properties remained constant until a degradation period of 26 weeks. Then they decreased continuously until they were completely lost at week 52. The molecular weight (M(w)) decreased steadily from 91,000 D in the case of the alpha-TCP composite and 78,000 D and 85,000 D in the case of the GB9N or GB14N composites, respectively, to about 10,000 D at week 78. It was concluded that the composites show adequate mechanical properties in the range of cancellous bone and a suitable degradation characteristic to be used as bone substitute materials.

Animals↗

Deproteinized cancellous bovine bone (Bio-Oss) as bone substitute for sinus floor elevation. A retrospective, histomorphometrical study of five cases.

OBJECTIVES: To study in detail the performance of deproteinized cancellous bovine bone (DPBB, Bio-Osso) granules as a bone substitute, a histomorphometric was performed on five patients treated with DPBB for reconstruction of the severely atrophic maxilla. MATERIAL AND METHODS: DPBB was used as mixture with autogenous bone particles, in concentrations that increased from 20% to 100% DPBB, with the time of healing increasing accordingly from 5 to 8 months. A total of 20 vertical biopsies was taken at the time of fixture installation and used for histomorphometry as undecalcified Goldner stained sections. RESULTS: The results show that in all cases, the DPBB granules had been interconnected by bridges of vital newly formed bone. The volume of bone in the grafted area correlated inversely with the concentration of DPBB grafted, and varied between 37% and 23%. However, the total volume of mineralized material (bone plus DPI3B granules) remained within the same range in all five patients (between 53% and 59%). The high values for osteoid and resorption surface, and the presence of tartrate-resistant acid phosphatase-positive multinucleated osteoclasts in resorption lacunae, indicated that bone remodeling was very active in all grafts. Osteoclasts were also observed in shallow resorption pits on DPBB surfaces. The percentage DPBB surface in contact with bone remained stable at about 35% and could not be related to the proportion of DPBB grafted. CONCLUSION: Although the number of patients examined was limited, the data suggest that deproteinized cancellous bovine bone, preferably combined with autogenous bone particles, is a suitable material for sinus floor elevation in the severely atrophic human maxilla.

Aged↗

Biomechanical behavior of hydroxyapatite as bone substitute material in a loaded implant model. On the surface strain measurement and the maximum compression strength determination of material crash.

Many investigators have advocated that hydroxyapatite ceramics may be extremely prospective bone substitute material mainly through evidence with its given biocompatibility towards bone and demonstrated continuity between living bone and hydroxyapatite. Its mechanical brittleness and strength, however, have been the most serious considerations. In this paper, a study has been performed using animal experiments, massive hydroxyapatite ceramics are implanted into rectangular bone defect created by operation at the load-bearing area in close contact with tibia plateau. The changes of surface strain at the hydroxyapatite implantation of retrieved tibiae are measured at any week after implantation when the compressive stress were applied in the direction of long axis in order to clarify the mechanical behaviors of hydroxyapatite living bone complex. The compression strength of hydroxyapatite implant crash was also determined at each week after implantation. Results revealed that the hydroxyapatite living bone complex has been proven to have sufficient flexibility such that it shows no hysteresis in stress versus strain relationship up to 200 kg of applied load. The strain pattern on the surface of hydroxyapatite implant develops quite similar to that of natural cortical bone. The material crash of hydroxyapatite implant tolerates up to 500 kg of load at 52 weeks after implantation. These characteristics suggest that it achieves normal skeletal function in the points of biomechanical properties in vivo.

Animals↗

Comparative study of biphasic calcium phosphate ceramics impregnated with rhBMP-2 as bone substitutes.

We investigated pellet-shaped implants prepared from biphasic calcium phosphate (BCP) ceramics with five different ratios of hydroxyapatite (HAP) to beta-tricalcium phosphate (beta-TCP) to evaluate these ceramics as bone substitutes. BCP ceramics impregnated with different doses of recombinant human bone morphogenetic protein 2 (rhBMP-2) (1, 5, and 10 microg) were used for experimental purposes and ceramics without rhBMP-2 were used for control. The pellets were implanted under the pericranium in adult Wistar male rats and were harvested 8 weeks after implantation. The retrieved pellets were then examined radiologically, histologically, and histomorphometrically. The results revealed that the pellets treated with rhBMP-2 exhibited new bone and bone marrow, whereas control pellets produced fibrous connective tissues. The formation of new bone induced by rhBMP-2 was dose dependent. The extent of bone and bone marrow formation and the degree of resorption of the ceramic particles were significantly higher in the pellets composed of 25% HAP-75% TCP. In this study, bioresorption of the ceramic produced favorable conditions for rhBMP-2-induced bone formation.

Adsorption↗

Continuous-release or burst-release of the antimicrobial peptide human lactoferrin 1-11 (hLF1-11) from calcium phosphate bone substitutes.

OBJECTIVES: In order to identify possible drug delivery systems against resistant bone infection, we determined the release of the antimicrobial peptide (AMP) human lactoferrin 1-11 (hLF1-11) from commercially available bone substitutes. METHODS: We combined six calcium phosphate cements and six granule-types with 5 mg/g hLF1-11 and measured its availability and release in vitro from cements (7 days) and granules (3 days). The integrity and antimicrobial activity of the hLF1-11 that was released during the first 24 h were measured, using mass spectrometry, and a killing assay on methicillin-resistant Staphylococcus aureus (MRSA). RESULTS: Most of the cements showed burst release followed by low-level continuous release, whereas the coated granules showed high burst release for 24 h. After release the peptide was active (in nine of 12 materials) and intact. CONCLUSIONS: Different release profiles may be obtained by choosing the appropriate carrier, which supports the feasibility of biodegradable carriers releasing AMPs against resistant infections.

Biodegradation, Environmental↗

Testing bone substitutes in a small animal model of revision arthroplasty.

This study evaluated a modification of the rat-pin model to enable testing of bone substitute materials. The model was characterized using the ceramic, beta-tricalcium phosphate (betaTCP) as a filler. A 1 mm wide, 3.6 mm deep defect was created around a stainless steel (SS) implant in the proximal tibia of a rat. This defect was filled with a ceramic powder. Large particles (90-312microm) of betaTCP were mixed with Gelfoam to form a paste which was then molded around the proximal end of either an uncoated SS pin or a pin coated with hydroxyapatite (HA). The pin with its ceramic collar was then implanted into the proximal tibia of 16 male Sprague Dawley rats. Two animals with coated implants and two with uncoated implants were sacrificed at 3, 6, 14 and 26 weeks. Longitudinal sections of each tibia were stained with toluidine blue and labeled for tartrate resistant acid phosphatase (TRAP). There was initial fibrous tissue interposition around the implants which was completely remodeled around the HA coated pins but which persisted in apposition to the SS pins. The remodeling process peaked at 3 weeks around the HA coated pins and at 6 weeks around the uncoated implants. There was little remodeling around either implant by 26 weeks. There was considerable residual betaTCP present which was well tolerated as the particles were often encased in bone. The model has several characteristics of revision arthroplasty and the results demonstrate the suitability of this model for testing bone substitutes.

Journal Article↗

Biomimetic PMMA-based bone substitutes: a comparative in vitro evaluation of the effects of pulsed electromagnetic field exposure.

Pulsed electromagnetic fields (PEMFs) are known to be effective in the stimulation of cultured osteoblasts and in vivo healing of delayed and nonunion fractures. In the present in vitro study the effects of PEMFs on osteoblastic cell cultures (MG63 human osteoblast-like cells) grown in the presence of poly-methylmethacrylate (PMMA) and of a biomimetic bone substitute made of a PMMA matrix added with alfa-tricalcium phosphate (PMMA+alpha-TCP) were evaluated, to assess the biological response at the cell-biomaterial interaction. Cultures were stimulated with PEMFs (75 Hz, 2.3 mT, 1.3-ms pulse duration) 12 h/day for 3 days and evaluations (MTT, ALP, OC, PICP, TGFbeta-1, IL-6) were performed at 3 and 6 days. PMMA had a negative effect on osteoblasts, whereas PMMA+alpha-TCP enhanced production of ALP, PICP, OC and TGFbeta-1, and reduced IL-6 levels. Cells responded positively to PEMF stimulation even when cultured with a poorly biocompatible material, such as PMMA. This effect was more evident in the presence of PMMA+alpha-TCP (further improvement in proliferation and synthetic activity) both at 3 and at 6 days. The properties of PMMA+alpha-TCP look promising, and the present results support the use of PEMFs to improve tissue response to biomaterials implanted as bone substitutes.

Bone Substitutes↗

Beta-tricalcium phosphate ceramic as a bone substitute in orthopaedic surgery.

Synthetic beta-tricalcium phosphate ceramic (Biosorb, S.B.M., Lourdes, France) was used as a bone substitute in 110 patients whose mean age was 48 (14-83) years. There was a wide spectrum of indications, and the material was used alone in 86, combined with a corticocancellous bone autograft in 22, and combined with a corticocancellous allograft in two. The patients were assessed clinically and radiographically. Incorporation of the ceramic was excellent in 30, good in 51, and fair in 26. There were no poor results. In three patients the implanted material was removed because of infection. Incorporation was good or excellent in about 75% of all 110 patients. We consider beta-tricalcium phosphate ceramic to be the substitute of choice for medium-sized bone defects.

Adolescent↗