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Biomedical subjects

G Daculsi

Publications and source records attributed to G Daculsi.

At least 91 records · Page 5Linked to original sources

[Morphometric study of the wall of the caudal vena cava after the implantation of a filter in the sheep].

The long stabilizers of the VT-LGM filter rest on large areas of the vascular wall. The morphometric study of the layers of the vascular wall, after implantation of filter in 15 ewes, divided into 3 groups of follow-up (2, 4 or 8 weeks) of 5 animals, was made at 3 levels opposite the filter and 1 level outside of it. Changes are seen for all the layers. Filter produces intimal hyperplasia. Stabilizers are quickly isolated from the blood flow. The thickness of the intimal hyperplasia grows for 4 weeks. It is more important at the base of the filter than at its head. There is also hyperplasia of the media with no change according to the follow-up or the level opposite the filter. The adventitia becomes thinner without variation of time or level of the filter. Smooth muscle cells of the adventitia become less frequent and smaller. Their density in front of the stabilizers, is the smallest by 4 week follow-up and remains the same along the stabilizer. The full thickness of the wall is bigger opposite the stabilizers than between them. The filter produces changes that are limited in time and in space. The most important changes are seen at 4 weeks after insertion and opposite the stabilizers.

Animals↗

Leukemia inhibitory factor and oncostatin M influence the mineral phases formed in a murine heterotopic calcification model: a Fourier transform-infrared microspectroscopic study.

The study of bone mineralization processes is of considerable interest in understanding bone diseases and developing new therapies for skeletal disorders, particularly since bone homeostasis requires numerous cell types and a large cytokine network. Cell culture models of mineralization have often been used to study the cellular mechanisms of mineralization, but few data have been reported concerning the influence of extracellular matrix components and cytokines on the physicochemical properties of mineral. The purpose of this study was to analyze the effects of two cytokines, leukemia inhibitory factor (LIF) and oncostatin M (OSM), involved in bone metabolism on the physicochemical properties of bone mineral formed in a murine in vivo mineralization model. Murine bone marrow cells implanted under the kidney capsule in the presence or absence of cytokines led to heterotopic ossicle formation. A scanning electron microscopic microprobe revealed that heterotopic calcification had a lower (approximately 20%) Ca/P ratio after cytokine treatment as compared with the control without cytokine. Transmission electron microscopic analysis of cytokine-treated ossicles showed numerous areas with low mineral density, whereas electron diffraction pattern revealed an apatitic phase. These areas were not observed in the absence of cytokine. Moreover, Fourier transform-infrared microspectroscopy showed at the molecular level that the presence of either cytokine induced many microscopic areas in which short-range order organization, such as incorporation of carbonate and crystallinity/maturity of ossicle mineral, were modified. LIF and OSM influenced mineral phase formation in the present model and may thus be key protagonists in bone mineral development and skeletal diseases.

Animals↗

Human growth hormone locally released in bone sites by calcium-phosphate biomaterial stimulates ceramic bone substitution without systemic effects: a rabbit study.

Calcium-phosphate bone replacement biomaterial has been used as a drug carrier for therapeutic agents. This study investigated the efficacy of local administration of human growth hormone (hGH) by macroporous biphasic calcium phosphate (MBCP) implants in improving the bone substitution qualities of ceramics. hGH release from MBCP implants loaded with 1 microg of hGH was rapid during the first 48 h and then sustained for a total of 9 days. Immunolocalization of hGH in vitro and in vivo by transmission electron microscopy showed its presence inside the material, indicating that it was able to penetrate within the porosity of the ceramic during the adsorption process. MBCP cylinders (6 x 6 mm) were loaded with 0.1, 1, and 10 microg of hGH and implanted into rabbit femurs (n = 40). The effects of locally released hGH on bone ingrowth and ceramic resorption were evaluated by scanning electron microscopy and image analysis. The results indicated that hGH increased bone ingrowth (+65%) and ceramic resorption (+140%) significantly in comparison with control implants and that the increase was dose dependent. Biochemical parameters monitored in rabbit plasma and urine, as well as the absence of any significant difference between contralateral implants and the control, indicated that hGH did not produce detectable systemic effects. Thus, the use of MBCP appears to be effective for local delivery of hGH, resulting in improved bone substitution.

Animals↗

Heterotopic implantation of mouse bone-marrow cells: an in vivo model allowing analysis of mineral phases during mineralization processes.

Heterotopic calcification induced after implantation of bone-marrow cells under the murine kidney capsule was used to study the mineral phases occurring during the mineralization process. Ossicles were found to contain numerous osteoblastic cells that produced an organic matrix closely associated with active hematopoietic tissue. During implantation of bone marrow, needle-shaped microcrystals were progressively deposited on collagen fibers. The mineral formed in the heterotopic calcification consisted mainly of calcium phosphate. The distribution and density of the microcrystals were heterogeneous after 6 weeks of implantation but became homogeneous and well-crystallized after 10 weeks. The Fourier transform infrared microspectroscopy provided important spatial data on the nature of the mineral formed and the changes in the mineral environment. Similarities were noted between young bone (bone callus) and 6-week heterotopic ossicles, and between adult bone and 10- or 12-week heterotopic ossicles. The study demonstrated that murine heterotopic calcification under the renal capsule can be a very useful model for studying bone apatite formation during the mineralization process.

Animals↗

Osteoclastic resorption of biphasic calcium phosphate ceramic in vitro.

Neonatal rabbit bone cells were cultured for 1 and 4 days on biphasic calcium phosphate ceramic specimens to study the osteoclastic resorption of the ceramic. Scanning electron microscopic studies after removal of stromal cells with pronase E and EDTA revealed many osteoclast-associated lacunae on the ceramic surface. The degraded crystals inside the lacunae appeared to have been dissolved by acids. Cellular degradation of the ceramic was clearly due to the extracellular process characteristic of osteoclastic resorption.

Animals↗

LPS increases biomaterial degradation by human monocytes in vitro.

Different cell lines are involved during an immunological reaction, principally lymphocytes and monocytes. Monocyte/macrophage cells, which are among the first to appear in wound-healing and infection sites, are largely implicated in phagocytosis and could be involved in calcium-phosphate degradation. Their role in these processes may relate to cytokine secretions and/or their sensitivity to certain cytokines. We tested the behavior of human monocytes placed on the surface of biphasic calcium-phosphate (BCP) tablets in the presence of two lipopolysaccharide (LPS) concentrations. After short-term culture (48 h), cytokine release (IL-1beta, IL-6) was measured by ELISA, and morphological cell events and biomaterial degradation were observed in scanning electron microscopy. BCP surface pits were noted near cells stimulated by 0.5 microg/mL LPS but were not apparent with 10 microg/mL LPS. The number of lacunae on BCP was increased after LPS treatment of human monocytes. An upmodulation of IL-1beta and IL-6 (in culture medium) released by LPS-activated human monocytes was observed, indicating good cell stimulation. This study demonstrates that LPS-activated human monocytes can degrade the surface of calcium-phosphate ceramic and confirms the role of human monocytes in biomaterial degradation.

Biocompatible Materials↗

Apatite as carrier for growth hormone: in vitro characterization of loading and release.

Previous studies concerning bone drug delivery systems have provided little data about the amount of drug loaded, one of the essential factors for determining the dose/effect relationship. To investigate this factor, an adsorption method involving a therapeutic agent was tested in vitro on an apatitic calcium phosphate (AP). One milligram of human growth hormone (hGH) was deposited onto 0.1, 0.15, and 0.2 g of AP powder over a period of 24 h at 37 degrees C. The amount of hGH loaded was determined by subtracting the dose recovered from that applied on AP. The results show that 1 g of AP absorbed 9.48 mg of hGH. From 0.1 and 0.15 g of hGH-loaded AP, hGH was released in vitro for 2 and 3 weeks, respectively, with a 50% time release (T1/2) at 30 h and 72 h, respectively, for the two quantities. The amount of drug loaded and the determined release kinetics were compatible with the action pattern of hGH, indicating that hGH-loaded calcium phosphate supports are suitable for bone-growth promotion.

Apatites↗

Association of human growth hormone and calcium phosphate by dynamic compaction: in vitro biocompatibility and bioactivity.

The association of therapeutic agents with biomaterials has been achieved through various techniques, such as coating of the ceramic block surface or drug incorporation into ceramics. The dynamic compaction method recently was developed to consolidate drug-loaded calcium phosphate powder without a sintering step. In the present work, human recombinant growth hormone was loaded on biphasic calcium phosphate powder and consolidated by a specific process of cold sintering (dynamic compaction). Analyses of the biocompatibility of compacted pellets (mouse L929 fibroblastic cell culture) and the bioactivity of the drugs released by them (growth hormone bioassay) were performed. This report demonstrates the biocompatibility of the compacts prepared by dynamic compaction. L929 cell proliferation was maintained and the capacity to secrete fibronectin was conserved in the presence of compacted materials. Comparison of released growth hormone integrity, revealed by radioimmunoassay and eluted stain bioassay, has shown that the biological activity of growth hormone was totally preserved after dynamic compaction. However, 35% of loaded growth hormone was not released in our experimental conditions, probably because of the inaccessibility of growth hormone within the granulated compacts. Dynamic compaction shows good potential for the production of biomaterials capable of releasing therapeutic agents in situ.

Animals↗

Upmodulation of multinucleated cell formation in long-term human bone marrow cultures by leukaemia inhibitory factor (LIF).

Inflammatory processes are mediated by many cellular events involving different cell types (leukocytes, monocytes, stromal cells, etc.). Numerous soluble mediators regulate these reactions, including leukaemia inhibitory factor (LIF), a cytokine which may play an important role in inducing acute-phase protein synthesis by hepatocytes during inflammation. This study was designed to determine the effects of LIF on the human monocyte/macrophage lineage and provide a better definition of its behaviour during systemic inflammation. In-vitro exposure of human long-term bone marrow cultures to recombinant human LIF significantly increased (about two-fold) the number of multinucleated cells (MNC) formed after three weeks of culture. These LIF-induced MNC expressed tartrate-resistant acid phosphatase, and LIF increased this intracellular activity by about 50%. MNC displayed phagocytotic activity but were unable to degrade sperm whale dentin or respond to human calcitonin. They did not possess the main characteristics of osteoclasts and were in fact macrophage polykaryons. Our results demonstrate for the first time that LIF can induce macrophage polykaryon formation from human bone marrow culture, suggesting that this factor not only produces leukocytes but also has a direct influence on the monocyte/macrophage lineage.

Acid Phosphatase↗

Adaptive crystal formation in normal and pathological calcifications in synthetic calcium phosphate and related biomaterials.

Mineralization and crystal deposition are natural phenomena widely distributed in biological systems from protozoa to mammals. In mammals, normal and pathological calcifications are observed in bones, teeth, and soft tissues or cartilage. We review studies on the adaptive apatite crystal formation in enamel compared with those in other calcified tissues (e.g., dentin, bone, and fish enameloids) and in pathological calcifications, demonstrating the adaptation of these crystals (in terms of crystallinity and orientation) to specific tissues that vary in functions or vary in normal or diseased conditions. The roles of minor elements, such as carbonate, magnesium, fluoride, hydrogen phosphate, pyrophosphate, and strontium ions, on the formation and transformation of biologically relevant calcium phosphates are summarized. Another adaptative process of crystals in biology concerns the recent development of calcium phosphate ceramics and other related biomaterials for bone graft. Bone graft materials are available as alternatives to autogeneous bone for repair, substitution, or augmentation. This paper discusses the adaptive crystal formation in mineralized tissues induced by calcium phosphate and related bone graft biomaterials during bone repair.

Animals↗

Dynamic compaction: a new process to compact therapeutic agent-loaded calcium phosphates.

The sintering stage in the classical process of preparing bone substitution materials prevents therapeutic agents from being loaded into calcium phosphate powder. However, dynamic compaction, a new process, requires no external heat, allowing the therapeutic agent to be incorporated into ceramics. This report presents the results of an in vitro study of therapeutic agents associated with calcium phosphate powder and involving this new process. A mixture of vancomycin lyophilized powder (0.15 g) and biphasic calcium phosphate (BCP) powder (1.85 g) was compacted. In addition, 2 ml of human growth hormone (1 mg ml-1) were associated with BCP powder by physical adsorption on bead surfaces before compaction. Detection by monoclonal antibodies and sodium dodecyl sulphate polyacrylamide gel electrophoresis demonstrated the structural integrity of the two therapeutic agents after consolidation. This new compaction process should be useful in developing ceramics that contain a therapeutic agent.

Adsorption↗

Osteoclastic resorption of calcium phosphate ceramics with different hydroxyapatite/beta-tricalcium phosphate ratios.

To study the influence of calcium phosphate ceramic solubility on osteoclastic resorption, neonatal rabbit bone cells were cultured for 2 days on hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP) and two types of biphasic calcium phosphate (BCP) with HA/beta-TCP ratios of 25/75 and 75/25. Solubility was regulated by varying the ratio of less-soluble HA and more-soluble beta-TCP. After removal of stromal cells by pronase E treatment, ceramic surfaces were observed by scanning electron microscopy. Osteoclasts resorbed BCP most extensively, with an HA/beta-TCP ratio of 25/75, producing typical lobulated, zig-zag track-like resorption lacunae. On pure beta-TCP, which had the highest solubility in acid, osteoclasts formed smaller discontinuous island-like lacunae. The resorption pattern may have been modified by the large number of calcium ions released into the acidic microenvironment at the osteoclast-ceramic interface. No resorption lacunae were found on the other specimens. The extent of osteoclastic resorption of calcium phosphate ceramics might, to a certain degree, be proportional to solubility, although this was not the case when solubility was very high. It would appear that ceramic solubility influences osteoclast resorption activity.

Animals↗

Fourier-transform infrared spectroscopy study of an organic-mineral composite for bone and dental substitute materials.

A new injectable biomaterial for bone and dental surgery is a composite consisting of a polymer as a matrix and bioactive calcium phosphate (CaP) ceramics as fillers. The stability of the polymer is essential in the production of a ready-to-use injectable sterilized biomaterial. The purpose of this study was to detect possible polymer degradation which may have been caused by the interaction with the fillers using Fourier transform infrared spectroscopy. Composites containing CaP fillers (biphasic calcium phosphate, hydroxyapatite and peroxidized hydroxyapatite) and polymer (hydroxypropyl methyl cellulose) were prepared. To investigate the properties of the polymer, the inorganic and organic phases of the composite were separated using several extraction methods. The difficulty in separating the organic (polymer) from the mineral (CaP fillers) phases in the composite investigated in this study suggested the presence of strong interactions between the two phases. Spectra of extracted polymers showed new absorption bands of low intensities and indications that some chemical modifications of the original polymers have occurred. Results also indicated that the filler composition has an effect on the integrity of the polymer.

Journal Article↗

Ovine anterior cruciate ligament reconstruction using a synthetic prosthesis and a collagen inductor.

The evaluation of composite anterior cruciate ligament prostheses, the union of an inductor of collagen and a synthetic fibre, are described. They were implanted in 10 sheep models for six months. None of the ligaments were broken, and the dynamic radiography was stable during this time. After harvesting them, a histologic study was performed on the intra-osseou and intra-articular portion, and on the synovial tissue. Biocompatibility was excellent. An osseous anchorage was found in 50% of cases and a fibrous ingrowth with well oriented fibres in each case. The results of this study show that the matrix has only partially played its role, because there was no improvement of fibrous ingrowth compared with other studies. However this fact corresponds to results at only 6 months and it is necessary to identify the type of collagen.

Journal Article↗

Three-dimensional defects in hydroxyapatite of biological interest.

Hydroxyapatite (HA) ceramics are widely used as bone substitutes in the repair of bony defects. These ceramics sometimes differ in their sintering temperatures. High resolution transmission electron microscopy was used to characterize HA ceramics sintered at different temperatures. A new type of defect was observed for the first time for ceramics prepared at 900 degrees C but not on those prepared at 1250 degrees C. This may cause a difference in their in vitro dissolution and in vivo performance.

Animals↗

Macroporous biphasic calcium phosphate ceramics: influence of five synthesis parameters on compressive strength.

Compressive strength measurements were conducted on 32 macroporous biphasic calcium phosphate (MBCP) samples to evaluate the influences and interactions of five synthesis factors: chemical composition, percentage of macropores, mean size of macropores, isostatic compaction pressure, and sintering temperature. These parameters were varied simultaneously between two limit levels. Experiments used a factorial design method (FDM) allowing optimization of the number of samples as well as statistical analysis of results. FDM showed that compressive strength, in a defined experimental area, can be described by a first-order polynomial equation in which the percentage of macroporosity and sintering temperature are the major influences. This study leads up to an isoresponse line diagram that will allow the manufacture of some classes of MBCP with fitted compressive strength.

Bone Substitutes↗

New preparation and microstructure of the EndoPatch elastin-collagen containing glycosaminoglycans.

Several improvements of the basic reaction between elastin-solubilized peptides and type I + III collagens are presented. They concern the prior incubation of the proteins in an adequate medium as well as the addition of heparan sulphate or hyaluronic acid (HA) and the consequent effects on the physical properties of yielded matrices. The addition of HA in small amounts enabled us to make membranes with good performances.

Biocompatible Materials↗

Biodegradation of synthetic biphasic calcium phosphate by human monocytes in vitro: a morphological study.

Biodegradation processes (both intra- and extracellular) occur immediately after implantation of calcium phosphate (CaP) ceramics. Monocytes and macrophages, among the first cells to appear in wound healing, are largely implicated in phagocytosis and may be involved in CaP degradation because of their sensitivity to secreted cytokines. We tested the behaviour of human monocytes placed on the surface of hydroxyapatite (HA) and biphasic calcium phosphate (BCP) tablets in the presence of vitamin D3 (VD3) and interferon gamma (INF gamma). After short-term culture (6 days), morphological events were observed in histological and scanning electron microscopy studies, and degradation lacunae were characterized. There were cell prints but no pits on the HA surface, but pits appeared near cells on the BCP surface. Preincubation of biomaterial in culture medium was essential. Variations in cell morphology were observed in different culture types. In the presence of VD3, degradation was greater than in the control, and cells were more polarized and rounded. With INF gamma, cells were extensively spread out on the sample surface, and the biomaterial seemed to be extracted from the surface by cells. Thus, monocytes are clearly influenced by soluble factors (vitamins, cytokines) and could be key cells in initiating the degradation of biomaterial.

Biodegradation, Environmental↗