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 Choukroun's 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 Choukroun's PRF in numerous implantology clinical applications.

Comparative study of manual and ultrasonic instrumentation of cementum surfaces: influence of lateral pressure.

The goal of periodontal treatments is to eliminate bacteria and their products without damaging cementum surfaces. Nonsurgical treatments are often limited by the inability of curettes to access the most apical zone of the pocket. While ultrasonic mini-inserts have been used for nearly 10 years now, their effect on dental tissues has not been tested. The purpose of the present study was to compare a new series of mini-inserts to Gracey curettes, which are the reference in nonsurgical treatments. Two experienced periodontists conducted root treatments on teeth destined for extraction using regular clinical criteria. One face of each root was instrumented using a Gracey curette, and the opposite face was instrumented using an ultrasonic mini-insert. After the instrumentation procedure, the teeth were prepared for examination by secondary electron (topographic features) and backscattered electron (organic and mineral composition) microscopy. Differences in surface composition between teeth treated by the two periodontists were noted and were related to the lateral pressure exerted. Calculus removal was less effective when strong lateral pressure was exerted using the ultrasonic mini-inserts, while more cementum was removed and more scratching occurred with both manual and ultrasonic instruments. In all cases, the ultrasonic mini-inserts allowed greater apical access. The new ultrasonic mini-inserts were as effective as manual curettes in eliminating plaque and calculus. The shape of the mini-inserts made them more effective in apical zones. The amount of damage to the cementum depended on the lateral pressure exerted by the periodontist.

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.

Effect of hydroxyapatite and beta-tricalcium phosphate nanoparticles on promonocytic U937 cells.

OBJECTIVES: Nanoparticles from implanted materials are reported as the main cause of implant failure. Monocytes are among the first cells to colonize the inflammatory site. We evaluated the biological effects of bone substitutes presented to U937 cells in vitro as micron- or nanometer-sized particles. METHODS: The HA (550 nm) and beta-TCP (550 nm) nanoparticles were incubated with U937 cells. Cell cycle modification, specific antigens expression, and the extent of cell death were determined. RESULTS: Firstly, by using the sulforhodamine B (SRB) test and the annexin V-FITC analysis by flow cytometry, our results provide evidence of the absence of cytotoxicity, and show that nanoparticles do not induce more apoptosis than microparticles in U937 cells. Secondly, although morphologic evidence of stimulation of U937-cells was found by confocal microscopy, neither bone substitute altered the distribution of the cells into different phases of the cell cycle (Kit Cycle Test Plus DNA). These results suggest that nanoparticles do not cause promonocyte maturation in macrophages. Thirdly, the flow cytometry results showed no differences in the expression of the adherence and activation markers. SIGNIFICANCE: The results suggest that nanoparticles do not promote the differentiation of promonocytic U937 cells into macrophages and do not induce an enhanced inflammatory response.

[Enamel micro-abrasion]. / 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 bond strength between simple foil mesh and laser-structured base retention brackets.

The aims of the current study were to evaluate the bond strength of a new metallic orthodontic bracket with a laser structured base (Discovery, Dentaurum, Ispringen, Germany), and its effects on the site of bond failure and on the behavior of the enamel after debonding. One hundred and twenty recently extracted human premolars were bonded with 1 of 2 types of mechanical interlock base metal brackets: a standard system with a simple foil mesh pad (Minitrim, Dentaurum) and the Discovery bracket. A resin-based, chemically activated bonding system, No-mix (Dentaurum), was used as the adhesive system in this trial. The teeth were immersed in normal saline solution at 37 degrees C for 7 days before debonding and were randomly assigned to different subgroups. A testing machine was used to evaluate tensile and fatigue bond strengths for both brackets. After debonding, the amount of residual adhesive on the bracket and enamel detachment were assessed according to the adhesive remnant index (ARI) and the enamel detachment index (EDI) with a scanning electron microscope and an energy dispersive X-ray spectrometer. The scores obtained from the ARI and the EDI showed that the laser structured base brackets had a significantly higher bond strength (mean +/- SD: 17.1 +/- 0.7 MPa) that was 2 times higher than that observed with the simple foil mesh brackets (mean +/- SD: 8.7 +/- 1.4 MPa) (P <.001). Bond failure with the laser structured base was at the enamel-adhesive interface with an ARI score of 3 in 80% of the teeth, and bond failure with the simple foil mesh base was at the bracket-adhesive interface with an ARI score of 0 in 75% of the teeth. A small area, with less than 10% of the enamel damaged (1 on the EDI) and 1.5 microm in thickness, was observed for both brackets. The laser structured base bracket's bond strength was double that of the simple foil mesh bracket but was equally safe and did not induce significant enamel detachment.