Guy Cathelineau

Apatite forming ability and cytocompatibility of pure and Zn-doped bioactive glasses

The use of bone grafts permits the filling of a bone defect without risk of virus transmission. In this work, pure bioactive glass (46S6) and zinc-doped bioactive glass (46S6Zn10) with 0.1 wt% zinc are used to elaborate highly bioactive materials by melting and rapid quenching. Cylinders of both types of glasses were soaked in a simulated body fluid (SBF) solution with the aim of determining the effect of zinc addition as a trace element on the chemical reactivity and bioactivity of glass. Several physico-chemical characterization methods such as x-ray diffraction, Fourier transform infrared spectroscopy and nuclear magnetic resonance methods, with particular focus on the latter, were chosen to investigate the fine structural behaviour of pure and Zn-doped bioactive glasses as a function of the soaking time of immersion in SBF. Inductively coupled plasma-optical emission spectroscopy (ICP-OES) was used to measure the concentrations of Ca and P ions in the SBF solution after different durations of immersion. The effect of the investigated samples on the proliferation rate of human osteoblast cells was assessed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, and tested on two different sizes of pure and zinc-doped glasses in powder form, with particle sizes that ranged between 40 to 63 µm and 500 to 600 µm. The obtained results showed the delay release of ions by Zn-doped glass (46S6Zn10) and the slower CaP deposition. Cytotoxicity and cell viability were affected by the particle size of the glass. The release rate of ions was found to influence the cell viability.

Aluminosilicates and biphasic HA-TCP composites: studies of properties for bony filling.

Aluminosilicate materials synthesized at room temperature present good mechanical properties. Hydroxyapatite, tricalcium phosphate or both offer a high biocompatibility in the biomedical field. In this work, we focused on the composites resulting from associations of these materials. The best compromise between porosity and biomechanical properties versus different parameters was determined. The in vitro behaviour of compounds in contact with the simulated body fluid (SBF) was studied and in vivo experiments in a rabbits thighbones were carried out. The inductively coupled plasma-optical emission spectroscopy (ICP-OES) method permitted us to study the eventual release of Al from composites to SBF and to evaluate the chemical stability of composites characterized by the succession of SiO(4) and AlO(4) tetrahedra. The kinetic biomineralization, the bioconsolidation and biological studies were made. The results obtained show the chemical stability of composites. In the bone-implant interface, the intimate links reveal the high quality of the biointegration and the bioconsolidation between composites and bony matrix. Histological studies confirm good bony bonding and highlight the total absence of inflammation or fibrous tissues.

In vitro effects of Choukroun's platelet-rich fibrin conditioned medium on 3 different cell lines implicated in dental implantology.

Objective: The aim of this work was to determine the relevance of Choukrouns platelet-rich fibrin (PRF) in dental implantology by determining the in vitro effects of soluble factors released by PRF clot. We used 3 different cell lines implicated in dental implantology: osteoblast, keratinocyte, and fibroblast. Methods: Cellular viability, cell proliferation, and gene expression were analyzed using PRF conditioned medium. Three different cells lines were used: SaOS2 (osteoblast), MRC5 (fibroblast), and KB (epithelial cell). Results: The sulforhodamine B assay showed a significant increase in cell number for the undiluted and 1:3 diluted conditioned medium after 24 and 48 hours. There was no effect for the 1:9 dilution. Cell cycle analysis by flow cytometry confirmed the viability test results. After 48 hours, PRF conditioned medium induced gene expression in osteoblasts. Expression of osteopontin and osteocalcin, late osteogenic markers, was observed using reverse transcriptase-polymerase chain reaction (RT-PCR). Conclusions: This study establishes a model to evaluate, in vitro, the effects of soluble growth factors released by PRF clot. Our work confirmed PRF is useful in stimulating tissue healing and bone regeneration. This work should recommend Choukrouns PRF in numerous implantology clinical applications.

Development of a three-dimensional model for rapid evaluation of bone substitutes in vitro: effect of the 45S5 bioglass.

The evaluation of innovative bone substitutes requires the development of an optimal model close to physiological conditions. An interesting alternative is the use of an immortalized cell line to construct multicellular spheroids, that is, three-dimensional (3D) cultures. In this study, a modified hanging drops method has resulted in the generation of spheroids with a well-established human fetal osteoblasts line (hFOB 1.19), and tests have been focused on the effect of 45S5 bioglass ionic dissolution products in comparison with two-dimensional (2D) cultures. Depending on cell culture type, quantitative analysis (cell proliferation, viability/cytotoxicity, and cellular cycle) and qualitative analysis (electron microscopy and genes expression) showed a differential effect. Cell proliferation was enhanced in 2D-conditioned cultures in accordance with literature data, but decreased in 3D cultures submitted to the same conditions, without change of gene expression patterns. The decrease of cell proliferation, observed in conditioned spheroids, appears to be in agreement with clinical observations showing the insufficiency of commercially available bioglasses for bone repairing within nonbearing sites, such as periodontal defects or small bone filling, in general. Therefore, we suggest that this model could be adapted to the screening of innovative bioactive materials by laboratory techniques already available and extended monitoring of their bioactivity.

Bioactivity behaviour of biodegradable material comprising bioactive glass

Biocomposite of bioactive glass (BG) with chitosan polymer (CH) is prepared by freeze-drying technique. Obtained material is investigated by using several physico-chemical methods. The XRD and FTIR show the interface bonding interactions between glass and polymer. The specific surface and porosity of biocomposite were determined. In vitro assays were employed to evaluate the effect of chitosan addition on the glass by studying the chemical reactivity and bioactivity of the BG and BG/CH biocomposite after soaking in a simulated body fluid (SBF). The obtained results show the formation of a bioactive hydroxycarbonate apatite (HCA) layer and highlight the bioactivity and the kinetics of chemical reactivity of bioactive glass, particularly after association with chitosan. The BG/CH biocomposite has excellent ability to form an apatite layer. Inductively coupled plasma-optical emission spectrometry (ICP-OES) highlights the negative effect of chitosan on the silicon release toward the SBF of bioactive glass when in vitro assays.

Le micro-sablage amélaire

Enamel conditioning (elimination of dental plaque and creation of an irregular surface) is an essential step before bonding of orthodontic brackets. The most popular procedure in our practice is bonding with resin which requires enamel etching in order to get enough shear bond strength. Many studies have tried to evaluate the effects of enamel bonding using the acid-etching procedure as well as the changes caused by detachment of brackets. Thanks to the development of other adhesives such as glass ionomer cements which chemically bind to the enamel, new enamel conditioning methods appeared, in particular sandblasting with aluminium oxide particles. This technique is a mechanical preparation of the tooth that avoids the harmful effects of acid products. By suitably choosing the parameters of sandblasting (pressure, time and quantity of powder), enamel loss is lower than with the acid-etch procedure and the surface of the enamel seems less affected. However the bond strength remains superior to the values required for treatment. The presented results indicate that enamel sandblasting can be considered as an alternative for the acid-etching technique currently used in orthodontic practice because it creates sufficient strength and respects enamel thickness better.

Comparison of the bony remodelling of two synthetic biomaterials: aragonite 55% and aragonite 55% with active substance

In this work, the in vivo behaviour of pure aragonite and vectabone, which is an association of aragonite and an active substance such as gentamicin, was studied to highlight the kinetic resorption of these two biomaterials with 55% of porosity destined for the filling or replacement of bony defects. The synthesis conditions and parameters we used permit us to obtain a biomaterial without a sintering stage. These conditions allow introducing of active substances at the first stage of the elaboration. In this work, the gentamycin antibiotic was associated with calcium carbonate (aragonite 55% with gentamycin) to deliver this active substance on the surgical site for local treatment. The tricalcium phosphate biomaterial was used as the control because of its high biocompatibility. The bony remodelling of these three biomaterials was studied by in vivo experiments. This study was ensured with neutron activation analysis (NAA). The resorption kinetic was elaborated and comparisons of the remodelling biomaterials CaCO(3) 55% and CaCO(3) 55% with gentamicin (vectabone) and tricalcium phosphate were carried out. The obtained results show that, 6 months after implantation, the mineral composition of vectabone and tricalcium phosphate becomes close to that of young bone. Twelve months after implantation, it becomes similar to that of mature bone.

In Vivo Mineral Composition Evolution of a Synthetic CaCO3 Aragonite Used as Biomaterial for Osseous Substitution

Abstract In orthopedic surgery, grafts are used as a substitute for defective bones. The development of synthetic biomaterials has been greatly successful. In this work, the in vivo behavior of synthetic aragonite calcium carbonate as a biomaterial was studied. This material porous at 44% was synthesized in our research group by double decomposition in an aqueous environment of salts of calcium and carbonate. Animal experimentation has been achieved on ovine spongy bony site. Biopsies have been carried out according to periods of 1, 3, 6, and 12 months after implantation. Samples were the subject of biologic assessments and in vivo mineral composition evolution. The analysis of trace and major elements (Ca, Sr, and P) of the biomaterial implanted has been achieved by the proton induced x‐rays emission (PIXE) method. Cartographies of every element of the biomaterial, of the interface (biomaterial–bone), and finally, of the bony matrix were established. Results obtained highlight the centripetal migrations of phosphorus and the strontium, from the physiological environment toward the aragonite CaCO3 versus decreasing time of the calcium concentration in the biomaterial. The elements migration kinetics is inhomogeneous in the biomaterial–bone interface area. These results have been correlated with histological studies.

Comparison of the Chemical Reactivity and Bioactivity of Two Bioactive Glasses 47S6 and 48S4

This work is focused on the bioactive glasses obtained by melting and rapid quenching. Two glasses with mineral composition of: 47% SiO2 - 26% CaO - 21% Na2O - 6% P2O5 and 48% SiO2 - 30% CaO - 18% Na2O - 4% P2O5 were investigated. The aim of this study was to establish the kinetics of HCAp layer formation “in vitro” and to control the adhesion and proliferation cells of the two glasses in contact with osseous cells. Obtained results permit to evaluate their chemical reactivity and their bioactivity after immersion in the SBF-K9. Ionic exchanges between biomaterials and SBF liquid during the “in vitro” experiments highlight the differences of the chemical reactivity and bioactivity of 47S6 and 48S4. The structural basis for the effect of cristallinity on the rates of HCA formation in vitro in favour of glasses was also established. The melt derived 47S6 and 48S4 glasses offer to surgeons new compositions with different bioactivity kinetic that bioglassÒ 45S6 and can be adaptable in some other bony pathology.

Physicochemical Reactivity after the In Vitro Assays of Synthetic and Natural Hydroxyapatite

Bioactive ceramics such as bioactive glasses, calcium carbonate and sintered hydroxyapatite are widely used in biomaterials field because of their high biocompatibility. In this study, natural hydroxyapatite (N-HA) and synthetic Hydroxyapatite (S-HA) were heat treated at 800°C and studied using “in vitro” experiments. Several physicochemical methods like: SRD, FTIR, SEM and ICP-OES were employed to evaluate the effects of the thermal treatment and to compare their behaviour after soaking in the Simulated Body Fluid SBF at different times. The formation of TCP has favours the formation of new phase. Obtained results show that the dissolution occurred more in N-HA than in S-HA and consequently the precipitation of new phosphate phase is more important in N-HA. This is due to the presence of Mg, Sr and Zn in N-HA with concentration higher to that in S-HA.