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Biomedical subjects

P Frayssinet

Publications and source records attributed to P Frayssinet.

At least 19 recordsLinked to original sources

[Cellular aspects of bone regeneration: role of bone marrow periostium].

Bone regeneration is only possible if stem cells give rise to progenitors of osteoblasts, chondroblasts or chondroidocytes. Stem cells and osteogenic progenitors were evidenced in bone marrow while only progenitors can be found in periosteum. Bone marrow stem cells did show an amazing plasticity and some cells of the bone surrounding tissues such as perivascular cells, adipocytes, muscle cells or even circulating cells are able to transdifferentiate in osteoblasts when submitted to an osteogenic environment. We have shown that the destruction of both bone marrow and periost impairs the bone healing. It indicates that the periost and bone marrow destruction removes the predetermined osteogenic cells and the informative factors able to induce the transdifferenciation of the cells contained in the peri-osseous tissues.

Bone Marrow↗

Effect of hydroxyapatite sintering temperature on intracellular ionic concentrations of monocytes: a TEM-cryo-X-ray microanalysis study.

Hydroxyapatite used as bone replacement can lead to particle release in the implantation site. These particles interact with monocytes, which are the first immune cells to colonize the implant and an inflammatory site. Thanks to cryo-X-ray microanalysis, we can observe cells in a state close to the physiological one and we have access to diffusible ions. We paid particular attention to the potassium-to-sodium ratio, which is one of the best viability criteria. We used this method to study the interaction between three hydroxyapatite particles treated at three different temperatures (not treated, treated at 600 degrees C and 1180 degrees C), and monocytes. In the culture condition, the hydroxyapatite treated at 1180 degrees C underwent the least dissolution. We demonstrate that monocytes were altered by the three hydroxyapatite particles. The hydroxyapatite particules treated at 600 degrees C were found to be more toxic.

Bone Substitutes↗

Adsorption and release of insulin-like growth factor-I on porous tricalcium phosphate implant.

In order to develop bone substitutes, the design of biomaterials like calcium phosphate ceramic loaded with bone growth factor are of great interest. However, it is necessary to control the amount of growth factor adsorbed onto ceramics and the kinetics of its release. Radiolabeling of insulin-like growth factor-I (IGF-I) with 125-iodine ([(125)I]-IGF-I) and its adsorption onto porous tricalcium phosphate (TCP) cylinders enabled us to establish the time-adsorption and time-release curves using various concentrations of IGF-I. The adsorption curve increased rapidly and then flattened out at 72 h; 90% of the maximum was already reached at 24 h; and 20% of the adsorbed IGF-I was released in water within 4 days. In human serum the release was faster at 82% within 4 days. In vivo evaluation on an animal model was then performed. Rabbits' bilateral femoral cylindrical bone defects were filled with the TCP cylinders, which were either carrying IGF-I or implanted alone as a control in each rabbit. Bone turnover and ceramic resorption were stimulated by IGF-I loaded TCP according to standard radiography, dual-energy X-ray absorptiometry, histology, and histomorphometry.

Adsorption↗

Osseointegration of composite calcium phosphate bioceramics.

The resistance of macroporous calcium phosphate ceramics to compressive strength generally is low and depends on, among other factors, porosity percentage and pore size. A compromise always is adopted between high porosity, required for a good integration, and mechanical strength, which increases with material density. We improved the strength of macroporous calcium phosphate ceramics of interconnected porosity by filling the pores with a highly soluble, self-setting calcium phosphate cement made of TCP and DCPD. Cylinders of the resulting material were implanted in sheep condyles and subjected to histological analysis after 20, 60, and 120 days. Microradiographs were made of the histological sections. The control material consisted of ceramic that had not been loaded with cement. Progressive ingrowth of bone into the ceramic pores occurred as the cement was degraded during the first implantation period. Marked degradation of the cement was apparent after 2 months, with fragmentation of the cement in most of the pores and the presence of bone tissue between the fragments. All the cement had been replaced by bone after 4 months. Some fragments of cement still were embedded in the newly formed bone. There was no significant difference between the integration of loaded and nonloaded ceramics. Filling the macroporous ceramic pores with a calcium phosphate cement significantly improved the mechanical strength of these ceramics without modifying their integration in the healing bone.

Animals↗

Accuracy and precision of in vivo bone mineral measurements in sheep using dual-energy X-ray absorptiometry.

We evaluated the precision and accuracy of in vivo measurements of spine bone mineral density (BMD) and bone mineral content (BMC) in five ewes using dual-energy X-ray absorptiometry (DXA, Lunar DPX-L). The short-term in vivo reproducibility expressed as the coefficient of variation (CV) varied from 0.9 to 1.6% for spine BMD and from 1 to 3.1% for spine BMC. The ex vivo measurements, performed in 20 cm of water to simulate soft tissue thickness, correlated closely with the in vivo measurements, yielding an r value of 0.98 and 0.97 for spine BMD and BMC, respectively. The accuracy was determined by comparing the total BMC of each vertebra measured in vivo with the corresponding ash weight. The correlation coefficient between the two measurements was r = 0.98, with an accuracy error of 5.6%. We concluded that the DXA allows a precise and accurate measurement of spine bone mineral in live ewes using the methodology designed for humans.

Absorptiometry, Photon↗

Integration of dense HA rods into cortical bone.

HA ceramics are daily used in human surgery for bone healing partly due to their ability to integrate into bone. They are generally used under a macroporous form. The behaviour of dense HA after implantation is not so well known. We implanted within cortical sheep femurs dense pure HA-ceramics cylinders for periods from 2 weeks to 18 months. The samples were then sectioned and examined using back-scattered and secondary SEM and the interface was analysed using EDS. Histomorphometry measurement was also performed using an image analysis device coupled to a light microscope. It appeared that the cylinders were in direct contact with immature bone after three weeks. The bone maturated within three months. The implant surface showed moderate signs of resorption and some grains were released from the surface. The resorption zone was only a few microm thick after 18 months. The bulk ceramic contained default zones of increased porosity. They can constitute fragile zone when located close to the surface in which the resorption rate is increased. We conclude that dense pure HA is poorly degraded when implanted in cortical bone. Degradation depends on the defaults found on the ceramic structure and the remodelling of bone surrounding the material.

Biocompatible Materials↗

Tissue reaction against a self-setting calcium phosphate cement set in bone or outside the organism.

Calcium phosphate cements are able to set in situ when injected into bone tissue. We evaluated the tissue reaction occurring when a DCPD-based calcium phosphate cement was either set within the bone or implanted when already set. The samples were implanted in rabbit condyles and examined histologically after 8 and 16 weeks. The relative bone surface, the fibrous capsule around the implants and the implant section surface were measured. Solid material seemed to be better tolerated than paste implants. More bone was found at the solid implant contact whatever the implantation time and the solid material degraded much less rapidly. In conclusion, the physico-chemical modification of the biological environment occurring during setting increases the foreign body reaction against the material.

Journal Article↗

[Osteolysis after total knee prosthesis].

PURPOSE OF THE STUDY: The aim of our work was to study X-rays showing osteolysis after 5 years and more in 122 prosthesis and to try and assess such complication, often described in the United States but seldom in Europ. MATERIAL: We are dealing here with 122 retaining posterior cruciate ligament, mostly cementless prothesis implanted between 1985 and 1992 84 chromium-cobalt prosthesis (PCA and Themis) implanted in 34 males and 88 females with an average age of 67 (45-81), 87,7 p. 100 had femoral cementless components and 70 p. 100 tibial cementless components. METHODS: All patients were examined and had X-rays at an average of 6,9 years. Specially considered were X-rays showing a possible osteolysis. We looked for possible complication (external laxity, anterior femoral dislocation and polyethylene wear), assessment of the mechanical axis and for clinical results (Hungerford score) RESULTS: Revisions: 19 arthroplasties were revised for PE wear tibial loosening metallosis or patella problems The postoperative score according to Hungerford was 84,5 p. 100 for PCA prosthese and 87 p.100 for the Themis. On the X-rays were only few osteolysis to be found: 9 cases (7,3 p. 100). For the PCA series: 3 femoral osteolysis, 1 tibial at 12 years, and one patellar osteolysis. For the Themis series: no femoral osteolysis, 3 tibial and one patellar osteolysis. Osteolysis are apparent on X-rays in profile for the femur and the patella, and in both profile and frontal X-rays for the tibia. Clinicaly 4 osteolysis really asymptomatic were not re-operated. 5 were revised: one 11 years later for femoral and tibial loosening, two for a patellar loosening, the other two patients had to be reoperated on for metallism (titanium's femoral component) and for those two instances osteolysis were discovered during the complication. DISCUSSION: Osteolysis after TKA appears unusual in our experience without bearing on frequency finded by american authors with a lesser follow-up (Engh 11,1 p. 100 after 4,5y, Peters 16 p. 100 after 2,9 y, Robinson 9,18 p. 100 after 4,6 y). American litteratur analysis shows that the important number of osteolysis is due to: - either to dual-metal using (Co-Cr component with Titanium screws for exemple), - or bad quality of polyethylene (compressed), - or a bad design of former prosthesis. CONCLUSION: Interface illness, linked to the production of wear debris, osteolysis after total knee arthroplasty is rarer than after a hip one, probably because size of debris is different, larger in knee than in hip. It is likely that the improvement of PE quality, design of prosthesis, as well as a better knowledge of osteolysis mechanism will allow to delay this complication wich is in a long term ineluctable.

Aged↗

[Engineering of osseous cells and bioartificial tissues].

The association of osteogenic stem cells to a synthetic carrier makes possible the elaboration of bioartificial tissue. Numerous phosphocalcic ceramics does not trigger a foreign body reaction when implanted in bone tissue and thus, a number of materials are available osteogenic stem cell carriers to replace the bone tissue. Several methods can be used to harvest these cells. Their multiplication in vitro can lead to the appearance of anomalies of their metabolism or their karyotype. The culture method also seems to have a major influence on their appearance. The presence of these anomalies could explain the variability of results in terms of bone extracellular matrix synthesis after cell reimplantation. The surgical technique used for the implantation is also of influence. A method suppressing the in vitro period has been developed to avoid any cell metabolism modification. This method allows for a very reproducible bone synthesis in ectopic site. The availability of human embryonic stem cells could help to develop cell graft techniques for bone reconstruction.

Biocompatible Materials↗

Degradation of hydroxylapatite, fluorapatite, and fluorhydroxyapatite coatings of dental implants in dogs.

Calcium phosphate coatings on dental implants enhance integration of the material. Resorption of the ceramic coatings has raised some concern about the behavior of the bone-implant interfaces after the coating disappearance. Substitution of the OH- ions by fluoride in the hydroxylapatite (HA) lattice makes the calcium phosphate more stable. We investigated the degradation rate of dental implants with 50- and 100-microm coatings of HA, fluorapatite (FA), or fluorhydroxylapatite (FHA). The implants were inserted in dog jaws and retrieved for histological analysis after 3, 6, and 12 months. The thickness of the calcium phosphate coatings was evaluated using an image analysis device. A relative resorption index and its standard deviation were studied. HA and FA coatings (even at 100-microm thickness) were almost totally degraded within the implantation period. In contrast, the FHA coatings did not show significant degradation during the same period. The standard deviation showed that the resorption process for FHA with thicknesses of 50 or 100 microm was the same. Such a difference was not observed between the 50- and 100-microm thick coatings of FA and HA. In conclusion, the FHA coatings showed good integration in the bone tissue and lasted much longer than classic calcium phosphate coatings.

Animals↗

Evaluation of human recombinant bone morphogenetic protein-2-loaded tricalcium phosphate implants in rabbits' bone defects.

Porous beta-tricalcium phosphate (betaTCP) has osteoconductive properties. The adsorption of human recombinant bone morphogenetic protein-2 (rhBMP-2) onto TCP could realize an osteoinductive bone substitute. We evaluated it on an animal model using dual-energy X-ray absorptiometry (DEXA) and solid-state 31P nuclear magnetic resonance (NMR) spectroscopy. BetaTCP cylinders loaded with rhBMP-2 were implanted into rabbits' femoral condyle bone defects, and betaTCP alone as control into the contralateral femur. We studied two different doses of rhBMP-2 (10 and 40 microg) on two groups of four animals. Evaluation consisted in radiography, histology, and histomorphometry, DEXA, and NMR spectroscopy using an original method of quantification. With both doses of rhBMP-2, we observed on radiographs an increase of trabecular bone around implants. Histology showed resorption of the ceramic, trabecular bone with osteoblasts and osteoid substance around the implants, and colonization inside the porous betaTCP by new bone formed. Histomorphometry showed that the osteoid surface (OS/BS) was greatest with the high dose of rhBMP-2. The difference was slight between the low dose of rhBMP-2 and control. DEXA showed a dose-dependent increase of bone mineral density of rhBMP-2-loaded betaTCP vs. control. NMR spectroscopy confirmed that the amount of new bone formed in betaTCP was greater when betaTCP carried rhBMP-2, and increased with the dose of rhBMP-2 used. We showed that betaTCP was a good matrix for rhBMP-2, which gave it osteoinductive properties in an orthotopic site, in a dose-dependent manner. Thus, such composite biomaterial seems to be of great interest in reconstructive bone surgery. Further studies are needed in clinical practice to determine optimal doses.

Absorptiometry, Photon↗

Biphasic synthetic bone substitute use in orthopaedic and trauma surgery: clinical, radiological and histological results.

Searching for an alternative to bone grafts in orthopedic and trauma surgery, two biphasic synthetic calcium phosphate ceramics BCP are made: the first, Eurocer 400, in granule form with a high interconnected porosity for void-filling, and the second, Eurocer 200, available in different shapes, with a good mechanical resistance in compression. Two hundred cases are reported with more than a six-month follow-up. The first 72 cases relating to hip arthroplasty revision surgery (29 involving acetabular and 43 femoral stem loosening) are exposed with some technical details. The next 71 cases concerning trauma and sequels are displayed with technical particulars. The remaining 57 other cold orthopedic indications are then enumerated. The very good biocompatibility of these ceramics is confirmed. Radiological incorporation is quickly seen in all the cases, faster with the filling substitute than with the second one, which, however, presents no mechanical failures when classical technical principles are respected. Finally, some histological studies are presented; the ceramics are progressively resorbed and bone reconstruction in and close to the substitutes is noticed. Therefore, the use of biophasic ceramics in almost all orthopedic and trauma surgery is recommended.

Journal Article↗

Silver methenamine staining for scanning electron microscopy of bone sections containing biomaterials.

Sections of tissue containing orthopedic materials are currently used to study the compatibility of those materials and to perform electron probe microanalysis at the material-tissue interface. Identification of the cells in contact with the material by Scanning electron microscopy (SEM) is of interest. We have developed a method for staining cells and tissue structures embedded in polymethyl methacrylate with silver methenamine once the sections have been obtained. Sections were prepared by grinding, and the silver methenamine was applied after oxidation with periodic acid. The procedure was carried out in a microwave oven. Backscatter SEM showed staining of the cell nucleus membrane, chromatin, the nuclear organizers, and the chromosomes of dividing cells. The cytoplasm and the cytoplasmic membrane were also stained. Collagen fibers of the extracellular matrix and the mineralized matrix of bone were labeled. Material particles in the macrophages were easily recognizable and Energy-Dispersive Spectrometer were not impaired by the presence of silver in the preparation.

Animals↗

Histopathology of a well-functioning hydroxyapatite-coated femoral prosthesis after 52 months.

A fully hydroxyapatite-coated femoral implant was retrieved during autopsy. This component, provided with a bipolar femoral head, had been inserted for a displaced fracture of the femoral neck 52 months before. Osseointegration of the implant was evident, without any formation of fibrous tissue :39.9% of the perimeter of the prosthesis at the level of its proximal third was interfaced with bone (62.8% at the mid-third and 65.2% at the distal third). Remodeling of bone had ensued. Deposition of bone was most prominent in the calcar zone, along the medial and lateral aspects and around the tip. Proximally, cortical porosity was found to be increased by 73%, whereas medullary bone porosity was increased by a factor of 2. Cell-mediated resorption of the coating was systematically present in these bone remodeling areas. The average thickness of the coating was respectively 10.8, 50.2 and 151.2 microns in the proximal, mid- and distal thirds of the implant. Formation of new bone was often coupled with resorption. No debris from the coating was found in the joint tissues or in the articulating surface of the polyethylene insert. These overall histopathological features support mechanical stability of the implant and active remodelling of bone along with focal removal of HA coating associated with osteoclastic activity. No side effects from coating degradation could be demonstrated.

Aged↗

Short-term implantation effects of a DCPD-based calcium phosphate cement.

Calcium phosphate cements can be handled in paste form and set in a wet medium after precipitation of calcium phosphate crystals in the implantation site. Depending on the products entering into the chemical reaction leading to the precipitation of calcium phosphates, different phases can be obtained with different mechanical properties, setting times and injectability. We tested a cement composed of a powder, containing beta-tricalcium phosphate (beta-TCP) and sodium pyrophosphate mixed with a solution of phosphoric and sulphuric acids. The cement set under a dicalcium phosphate dihydrate (DCPD)-based matrix containing beta-TCP particles. This was injected with a syringe into a defect drilled in rabbit condyles, the control being an identical defect left empty in the opposite condyle. The condyles were analysed histologically 2, 6 and 18 weeks after implantation. After injection into the bone defect the cement set and formed a porous calcium phosphate structure. Two different calcium phosphate phases with different solubility rates could be identified by scanning electron microscopy (SEM) observation. The less-soluble fragments could be degraded by cell phagocytosis in cell compartments of low pH or integrated in the newly formed bone matrix. The degradation rate of the material was relatively high but compatible with the ingrowth of bone trabeculae within the resorbing material. The ossification process was different from the creeping substitution occurring at the ceramic contact. Bone did not form directly at the cement surface following the differentiation of osteoblasts at the material surface. The trabeculae came to the material surface from the edges of the implantation site. Bone formation in the implantation site was significantly higher than in the control region during the first week of implantation. In conclusion, this material set in situ was well tolerated, inducing a mild foreign-body reaction, which did not impair its replacement by newly formed bone within a few weeks.

Animals↗

Histological integration of allogeneic cancellous bone tissue treated by supercritical CO2 implanted in sheep bones.

UNLABELLED: Different chemical or physical methods of bone processing have been developed to decrease the antigenicity of allogeneic bone which may delay or prevent graft integration. We have developed a method based on delipidation and deproteination of the bone with a supercritical fluid and hydrogen peroxide. Cylinders of cancellous allogeneic bone treated in this way were implanted for four weeks, four months or eight months in holes drilled in sheep condyles or tibial plateau. Histological sections were then processed and analysed qualitatively and quantitatively using an image analysis software coupled to a light microscope. Measurements were made of the trabecular bone surface (BS/TS), the relative osteoid surface (OS/BS), the active osteoid surface (OS/BS), active resorption surface (Oc.S/BS) and the relative surface of newly formed bone. After four weeks, the control cylinders (non-treated allogeneic bone) had been invaded by cellular tissue composed of lymphocytes and plasmocytes surrounding remnants of the donor bone marrow tissue. The processed cylinders showed osteoid apposition at the surface of their external trabeculae. The trabecular bone and osteoid surfaces were significantly higher in the processed bone sections than in the control bone sections. After four months, most of the control material had been osteolysed and replaced by connective tissue containing lymphocyte islets, while the processed materials showed a large amount of bone synthesized at the surface of implant trabeculae which appeared fragmented and disseminated within the newly formed bone. All the histomorphometric parameters measured were significantly different from those of the control. By eight months, most of the control material had been totally osteolysed with very little bone ingrown in the implantation site. Only one control implant had been integrated. The processed cylinders were difficult to discern from the bone in which they were implanted. The parameters measured on the processed cylinders were significantly higher than those measured on the control sections. IN CONCLUSION: the treatment applied to the bone enhanced allogeneic bone integration and could provide a new kind of tissue treatment for bone banking.

Animals↗

Reimplantation of growth plate chondrocyte cultures in central growth plate defects: Part I. Characterization of cultures.

Growth plate lesions or resections may cause severe growth arrest because of the bony bridge between the epiphysis and metaphysis. Actual treatments for epiphysiodesis include resecting the bone bar and setting an interpositional material. Growth plate cultures may provide the appropriate cartilage necessary to restore growth potential when implanted in a growth plate defect. The aim of this work was to determine certain cell culture parameters in order to optimize in vitro cultures to obtain abundantly mature and functional chondrocytes. We studied the manner in which enzymatic digestion, carried out by various enzymes, obtained chondrocytes. Treatment with trypsin (0.2%) during 30 minutes at 37 degrees C and then collagenase (200 U/mL) during 6 hours was chosen. Under these conditions, 40 +/- 16 10(6) chondrocytes per gram of growth plate were obtained, and cellular viability was 79 +/- 12%. The density of the cellular seeding, the nature of the culture substrate, and the culture medium composition were determined to optimize the growth of differentiated cells. Seeding at 20,000 or 30,000/cm2 on a type I substrate and Ham F-12 medium not supplemented with either glucose or growth factors was demonstrated to be the best choice for this purpose.

Animals↗

Reimplantation of growth plate chondrocyte cultures in central growth plate defects: Part II. Surgical experimentation in rabbits.

An original and reliable technique to culture growth plate chondrocytes was developed to obtain an abundant amount of mature and functional chondrocytes. Growth plates were provided from the epiphysis of 3-week-old rabbits. Isolation of the chondrocytes was optimized by the use of trypsin and collagenase. The culture was realized according to the following conditions: seeding at 20,000 or 30,000/cm2 on type I collagen substrate and in Ham F-12 medium without a supplementation of glucose or growth factors. After 7 days of culture, the implantation was to be carried out. Different implantation substrates were evaluated in vivo. Agar turned out to be the only substrate to provide strong and healthy chondrocytes 21 days after the grafting. Then implantation was tested on large iliac resections in rabbits to check whether an enchondral ossification occurred with the culture. Poor results were obtained because of an early disappearance of the cultured chondrocytes. In an other experimentation, the culture was implanted into surgically created defects in the growth plate area. In this case, the culture did produce an epiphysiodesis. However, the 6-week postoperative histological examination showed that the implant remained viable, continued to maintain a proteoglycanrich matrix, and began to organize in ordered columns of mature chondrocytes.

Animals↗