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Biomedical subjects

P Frayssinet

Publications and source records attributed to P Frayssinet.

36 records · Page 2Linked to original sources

[Coloration of cytologic thick sections containing biomaterials with silver methenamine. Usefulness for scanning electron microscopy].

UNLABELLED: Prostheses implanted in hard tissues cannot be processed for electron microscopic examination or microanalysis in the same way as those in other tissues. For these reasons, we have developed a method allowing light and electron microscopic studies as well as microanalysis of the interface between bone and biomaterials after the sections glycoproteins have been stained with silver methenamine. Silver can be evidenced by SEM in back scattered mode. MATERIALS AND METHODS: Tranverse sections of a human femur containing an HA-coated prostheses were obtained with a diamond saw and ground to a thickness of 50-100 microns. The sections were stained in a microwave oven using a 1% silver methenamine solution. They have been examined by back-scatter SEM operated at 25 KV. EDS has been performed on cellular inclusions and extracellular bone matrix. RESULTS: Type I and III collagen fibers, and reticulin fibers were stained. The mineralized matrix was heavily colored. At the cell level, the nuclear and cytoplasm membranes, the chromatin and ribosomes were shown. The characteristic peaks of the Ag spectrum are distinct from those of the elements used in orthopaedic biomaterials and did not impair their identification.

Animals↗

[Osseointegration of 2 different types of calcium phosphate materials: ceramics and ionic cements].

UNLABELLED: Different kinds of calcium phosphate biomaterials can be used as bone substitutes. Ceramics are constituted by HA or TCP grains linked by grain boundaries. Their porosity depends on the powder characteristics and the sintering temperature. It can be very low with a pore size inferior to one micron. The setting of calcium phosphate hydraulic cements results from the precipitation of a calcium phosphate phase different from the one in suspension in the paste. The strength of the cement is given by the entanglement of the growing mineral crystals. Calcium phosphate hydraulic cements and ceramics have very different physico-chemical characteristics. We have studied the histological integration of both kinds of material. The first material was constituted by macroporous ceramics composed of 75% HA and 25% beta-TCP, the cement was made of beta-TCP grains dispersed in a DCPD matrix. The sequence of events which leads to the ceramic integration is always the same: a/ ingrowth of a loose connective tissue; b/ osteoblast differentiation from fibroblast-like cells of the connective tissue in close proximity to the implant surface; c/ osteoid synthesis at the ceramic surface toward the pore center; d/ remodeling of the immature bone and the ceramic itself. The cement is differently integrated. The osteoblasts differentiate at some distance from the implant and there is a trabeculae ingrowth toward the material. CONCLUSIONS: The early stages of both materials osteointegration are different. The integration is centrifugal for ceramics and centripetal for the cement.

Animals↗

[Tartrate-resistant acid phosphatases (TRAP) and cytology of pseudosynovial membranes].

Pseudo-synovial membranes are found at the interface between bone and aseptically lost prostheses. They are supposed to be due to the release of material debris at the interface with bone. They are mostly composed of macrophages and giant cells, and some fibrocytes in the deep layer of the membrane. The different histological structures present in these membranes have been largely described. Very recently, Kadoya et al. have demonstrated that a lot of giant cells present in the pseudo-synovial membranes show a tartrate resistant acid phosphatase activity in the presence of more than 100 mM tartrate. They have also shown that some of these cells were responsible for bone degradation. We have studied the tartrate acid phosphatase activity of a series of 10 membranes containing various material debris: titanium alloy, stainless steel, polymethylmethacrylate, polyethylene. Many mono or multinucleated cells showed a TRAP activity in their lysosomes in the presence of a 400 mM tartrate concentration. In certain membranes, these cells showed a TRAP activity located in the nucleus. This study shows that the macrophage population is very heterogeneous and the difference between osteoclasts and giant cells may be unclear.

Acid Phosphatase↗

Osseointegration in cortical sheep bone of calcium phosphate implants evaluated by PIXE method and histology.

Osseointegration of porous calcium phosphate ceramics evolves in several stages once implanted. Histologic analysis has often been used to evaluate the mechanism of integration of this material. Histologic parameters can be completed by physical analysis to obtain a semiquantitative evaluation of the osseointegration process. The histologic observation of hydroxyapatite (HA)-ceramic-containing bone sections was associated with proton-induced X-ray emission (PIXE) analysis and the results obtained by both methods were compared. Porous HA-ceramic cylinders were implanted in cortical bone of sheep femurs for periods ranging from 2 to 36 weeks. Thick sections of the implant containing bone were made at the end of the implantation period. A scanning line with two proton impacts 0.5 mm apart was plotted from the edges of cortical bone across the implanted ceramic and the X-rays produced were determined. Calcium, phosphorus, zinc, strontium, and iron contents were measured. Following PIXE analysis, the sections were surface-stained and observed under a light microscope to define the osseointegration index. Two regions of the curves were identified for each element characterizing either the bone tissue or the ceramic. Zinc and strontium present in the bone tissue but absent from the ceramics appeared after the 8th and the 12th implantation weeks, respectively. Iron present in the implant decreased with time, and calcium and phosphorus contents tended to be the same at the end of the implantation period in both curve regions. Histologic observation showed that immature bone invaded the pores of the outer layer of the ceramic as early as 2 weeks after implantation. Ceramics were totally osseointegrated 20 weeks after implantation. Osseointegration was apparently still evolving as judged by the PIXE method when histologic integration was considered complete.

Animals↗

The influence of sintering temperature on the proliferation of fibroblastic cells in contact with HA-bioceramics.

HA-ceramics used in human surgery as osteoconductive surfaces show a great variety of characteristics. Certain characteristics such as grain size, porosity, and surface area, are controlled by the sintering temperature of the slurry. We grew L-929 fibroblast cells on HA-ceramic disks that had been sintered at different temperatures ranging from 850 degrees-1350 degrees C. The cell line growth rate was lower on ceramic disks than on the culture-grade polystyrene used as a negative control. Cell growth correlated with the ceramic sintering temperature although no significant difference in the cell adhesion to the different ceramics was shown. Growth rate on ceramics sintered at low temperatures (850 degrees and 950 degrees C) was negative whereas it was positive on disks sintered at higher temperatures. When the cells were separated from the disks by a polycarbonate membrane, the growth rate was negative on those membranes in contact with low-temperature sintered disks and positive on the high-temperature sintered disks. The calcium and phosphorus concentration in the culture medium in contact with ceramics sintered below 1050 degrees C decreased during the culture period. Ceramics sintered between 1100 degrees and 1250 degrees C brought about an increase in Ca and P concentrations while ceramics sintered at higher temperatures did not induce any changes. SEM examination of the 850 degrees and 1200 degrees C sintered ceramics showed that the 850 degrees C sintered ceramics consisted of small grains with pores between them and the 1200 degrees C sintered ceramics were made of larger grains without any visible pores, thereby decreasing the surface of material in contact with the culture medium. This difference in surface area was confirmed by the fact that the amount of albumin absorbed onto the ceramic was dependent on the sintering temperature. In conclusion, the modification of the culture medium brought about by high-surfaced ceramics could influence the growth of cells with which such ceramics come in contact.

Animals↗

Analysis of sections of implanted macroporous calcium phosphate bone substitutes by proton-induced X-emission method and energy-dispersive spectrometry.

The osseointegration of porous calcium phosphate ceramics once implanted evolves in several stages. The mechanism of integration of such material usually is evaluated by histologic analysis. The trace elements present in bone can be detected in the ceramic and help to provide a semiquantitative evaluation of osseointegration. Two different methods of microanalysis, energy-dispersive spectrometry (EDS) and proton induced x-emission (PIXE) were used in this study to determine the appearance of trace elements (Zn, Sr, and Fe) present in bone at the implantation site containing the ceramic. Porous HA-ceramic cylinders were implanted in the cortical bone of sheep femurs for periods ranging from 2 to 36 weeks. Thick sections of the implant-containing bone were made at the end of the implantation period. A scanning line with proton or electron impacts 0.5 mm apart was plotted from the edges of the cortical bone across the implanted ceramic and the resulting x-ray spectra were determined. Following EDS analysis, the sections were surface-stained, observed under a light microscope, and the pore volume occupied by bone tissue was measured. The spectra obtained by PIXE method showed two regions for each element characterising either the bone tissue or the ceramic. Zinc and strontium present in the bone tissue, but absent from the ceramic, appeared 8 and 12 weeks after implantation, respectively. The concentration of iron present in the implant decreased with time. EDS showed no significant level of either element in the bone or the ceramic. Histologic observation revealed that immature bone invaded the pores of the outer layer of the ceramic as early as 2 weeks after implantation. The ceramics were totally osseointegrated 20 weeks after implantation, although ceramic degradation continued for longer. In this experiment, the PIXE method was apparently sufficiently sensitive for monitoring the amount of trace element appearing in bone-implanted material.

Animals↗

Resorption kinetics of osseous substitute: natural coral and synthetic hydroxyapatite.

Coral and hydroxyapatite may be used as substitution biomaterials for bone grafts. In this work, we extracted the implants from the femora to study the kinetics of elementary mineral transformation of the osseous substitutes. The use of physical analysis methods such as PIXE (particle-induced X-ray emission) shows that coral and hydroxyapatite, after their implantation in vivo, reach a mineral composition comparable with that of bone. For the first time we have measured the concentration of mineral elements, at different time intervals after implantation, along a cross-section. The distribution according to mineral elements (Ca, P, Sr, Zn, Fe) in the implant, in the receiver site and also at the interface, showed that the kinetics of coral resorption was faster than that of hydroxyapatite; in the same way, the osseous attack was not global but, rather, centripetal.

Animals↗

Use of supercritical CO2 for bone delipidation.

Supercritical carbon dioxide was used for bone delipidation. It appeared that this technology is very efficient since supercritical CO2 is able to diffuse into microporous solids much better than liquids and that it has a good solvent capacity for lipids. This extraction is the ideal first step of any bone processing because microporosity of bone tissue becomes much more accessible, which may enhance osteoconduction once implanted. Moreover, it is safe since it involves no toxic chemical and is potentially usable with allografts as well as xenografts.

Animals↗

Two-year outcome of hydroxyapatite-coated prostheses. Two femoral prostheses retrieved at autopsy.

We performed a histological study of the bone-implant interface on 2 human femurs implanted with a hydroxyapatite-coated self-locking stem for 2 years. Extensive bone formation with no intervening fibrous tissue was noted around the entire circumference of the 2 prostheses. The newly-formed bone had 2 morphotypes: an alveolar disposition with a continuous contact between bone and hydroxyapatite, and a digitiform one where distinct bony trabeculae were in contact with the ceramic coating or with the bone marrow. Partial or even total resorption of the hydroxyapatite coating was clearly identified, these areas showing bone in contact with the metal.

Aged↗

Osseointegration of macroporous calcium phosphate ceramics having a different chemical composition.

Calcium phosphate macroporous ceramics are biocompatible for bone surgery. Their osseointegration is, however, sometimes very poor. To measure the effect of the calcium phosphate content of the ceramic on its osseointegration, macroporous ceramics, differing in their chemical composition, were implanted into sheep femurs. The ceramics were composed of different percentages of hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP). All other characteristics were the same. Results were assessed histologically with image analysis and showed significant differences in the amount of bone formed at the contact of the different ceramics. Ceramics containing beta-TCP induced better osseointegration than pure HA ceramics.

Animals↗

A study of structure and degradation of nonpolymeric biomaterials implanted in bone using reflected and transmitted light microscopy.

Orthopedic biomaterials currently are made of metal alloy coated with one or more thin layers of dense or porous ceramic or metal. Sections of these materials implanted in human bone were made without altering the implant or bone-implant interfaces. Bone containing an implant was fixed and then embedded in polymethylmethacrylate. Thick sections were made using a cooled, low speed diamond saw, then ground and polished. Some were stained by fuchsin-toluidine staining solution, others were acid etched to reveal the structure of the metal contained in the prosthesis. Observation by reflected and transmitted light microscopy revealed microstructure of the implant material as well as features of the surrounding tissues.

Alloys↗

[Histological analysis of the bone/prosthesis interface in man after implantation of a hip prosthesis coated with plasma-sprayed hydroxyapatite].

Thin coatings of calcium phosphate hydroxyapatite on metal alloys provide to these materials biological properties of calcium phosphates. We have analysed, using histological techniques or newly developed scanning electronic microscopy techniques, hip prostheses implanted into humans for periods from a few days up to twenty six months. The results of these analyses confirm the good osteointegration of these prostheses observed during clinical studies. Moreover, an active remodeling of the bone in contact with the ceramic-coating was observed. The coating was also concerned by the remodeling process and evolved once implanted.

Bone Matrix↗

[Hybrid substitute materials for bone. Initial results and perspectives].

Hybrid materials for use as bone substitutes are composed of a synthetic substance to which autologous osteogenic cells are bound. Identification of osteogenic stem cells among bone marrow stromal cells was followed by the demonstration that a host of growth factors, hormones, and cytokines have effects on cultured osteoblasts. Porous ceramics have been developed as vectors for osteogenic cells previously grown in vitro. Porous hydroxyapatite beads bearing stromal cells were used to fill experimental ulnar defects in dogs. The cells were first grown on flat supports then on the three-dimensional hydroxyapatite network. Invasion by histologically identifiable bone tissue was seen in the stroma cell-bearing implants but not in control implants without stromal cells. Materials containing osteogenic cells may prove extremely useful since bone substitutes including hydroxyapatite ceramics are implanted in sites where osteogenic cells are scarce.

Animals↗

Bonding of hydroxyapatite-coated femoral prostheses. Histopathology of specimens from four cases.

We examined specimens of hydroxyapatite-coated femoral prostheses from four patients who had died within nine months of implantation for fractured neck of femur. Histology showed newly formed immature bone overlying the hydroxyapatite coating with new trabeculae bridging to the endosteal bone layer. In the diaphysis, where there had been contact between the hydroxyapatite and the cortex, there was dense, firmly anchored bone with an haversian architecture. In other places the newly formed bone had a trabecular structure, containing bone marrow tissue with normal cellularity. It appeared that biological osseointegration had taken place.

Aged↗

Viral inactivation of human bone tissue using supercritical fluid extraction.

A new bone tissue process using supercritical carbon dioxide fluid extraction (SFE) has been evaluated for its ability to inactivate or eliminate viruses. Four viruses, human immunodeficiency virus type 1 (HIV-1), Sindbis virus, polio Sabin type I virus, and pseudorabies virus (PRV), were exposed to four different processing steps. In addition to supercritical CO2, hydrogen peroxide, sodium hydroxide, and ethanol treatments were evaluated. The mean cumulated reduction factors (log10) for the four viruses exposed to these four steps were > 14.2 for HIV-1, > 18.2 for Sindbis virus, > 24.4 for poliovirus, and > 17.6 for PRV. The mean reduction factors obtained by the supercritical fluid extraction alone were > 4.0, > 4.3, > 6.6, and > 4.0, respectively. These results demonstrate that the SFE process is effective in inactivating viruses on human femoral heads, and provides a level of inactivation similar to that obtained by traditional cleaning methods. It is proposed that CO2 SFE be incorporated as a routine step in the processing of bone allografts for transplantation either to replace or supplement existing procedures.

Carbon Dioxide↗