Complement Activation by Pulp Capping Materials Plays a Significant Role in Both Inflammatory and Pulp Stem Cells' Recruitment.

INTRODUCTION: The role of complement, especially through the C5a fragment, is well-known for the initiation of inflammation. Its involvement in regeneration has been shown more recently by the recruitment of mesenchymal stem cells. C5a can be produced locally by the pulp fibroblasts in response to injury or infection. This work aims to investigate the effect of different pulp capping biomaterials on complement activation and its possible influence on inflammatory and pulp stem cell recruitment. METHODS: Conditioned media were prepared from 3 pulp capping biomaterials: Biodentine (Septodont, Saint-Maur-des-Fosses, France), TheraCal (BISCO, Lançon De Provence, France), and Xeno III (Dentsply Sirona, Versaille, France). Injured pulp fibroblasts were cultured with these conditioned media to analyze C5a secretion using an enzyme-linked immunosorbent assay. Dental pulp stem cells (DPSCs) were isolated from human third molar explants by magnetic cell sorting with STRO-1 antibodies. The expression of C5a receptor on DPSCs and inflammatory (THP-1) cells was investigated by immunofluorescence. The migration of both DPSCs and THP-1 cells was studied in Boyden chambers. RESULTS: Pulp fibroblast production of C5a significantly increased when the cells were incubated with TheraCal- and Xeno III-conditioned media. The recruitment of cells involved in inflammation (THP-1 cells) was significantly reduced by Biodentine- and TheraCal-conditioned media, whereas the migration of DPSCs was reduced with TheraCal- and Xeno III-conditioned media but not with that of Biodentine. The involvement of C5a in cell recruitment is demonstrated with a C5a receptor-specific antagonist (W54011). CONCLUSIONS: After pulp injury, the pulp capping material affects complement activation and the balance between inflammation and regeneration through a differential recruitment of DPSCs or inflammatory cells.

Disc displacement without reduction: a retrospective study of a clinical diagnostic sign.

OBJECTIVE: The purpose of this retrospective study is to evaluate a clinical diagnostic sign for disc displacement without reduction (DDWR), the absence of additional condylar translation during opening compared with protrusion. METHOD: Thirty-eight electronic axiographic and magnetic resonance imaging (MRI) examinations of the TMJ were analyzed in order to compare the opening/protrusion ratio of condylar translation between non-painful DDWR and non-DDWR. RESULT: According to the Mann-Whitney U test, the opening/protrusion ratio in non-painful DDWR differs significantly from non-DDWR (p < 0.0001). DISCUSSION: Among non-painful DDWR, there is no additional condylar translation during opening in comparison with protrusion, and this is probably also the case for DDWR without limited opening, which is a subtype that has not been validated by the Diagnostic Criteria for Temporomandibular Disorders (DC/TMD). Comparative condylar palpation can analyze this sign, and therefore, further comparative investigations between MRI and clinical examination are needed to validate the corresponding clinical test.

Sources of dentin-pulp regeneration signals and their modulation by the local microenvironment.

Many aspects of dentin pulp tissue regeneration have been investigated, and it has been shown that dentin pulp has a high regeneration capacity. This seems to be because of the presence of progenitor cells and inductive regeneration signals from different origins. These signals can be liberated after the acidic dissolution of carious dentin as well as from pulp fibroblasts and endothelial cells in cases of traumatic injury. Thus, both carious lesions and pulp cells provide the required mediators for complete dentin-pulp regeneration including reparative dentin secretion, angiogenesis, and innervation. Additionally, all dentin pulp insults including carious "infection," traumatic injuries, application of restorative materials on the injured dentin pulp, and subsequent apoptosis are known activators of the complement system. This activation leads to the production of several biologically active fragments responsible for the vascular modifications and the attraction of immune cells to the inflammatory/injury site. Among these, C5a is involved in the recruitment of pulp progenitor cells, which express the C5a receptor. Thus, in addition to dentin and pulp cells, plasma should be considered as an additional source of regeneration signals.

In vitro microleakage of Biodentine as a dentin substitute compared to Fuji II LC in cervical lining restorations.

PURPOSE: 1) To evaluate the marginal sealing efficacy of Biodentine at the cervical margins of approximal cavities placed in molars; 2) to evaluate and compare the use of Biodentine in combination with resin-based adhesives and a resin composite, compared with a resin-modified glass-ionomer cement (Fuji II LC). MATERIALS AND METHODS: Sixty approximal cavities were prepared on mesial and distal surfaces of 30 extracted human third molars. The teeth were randomly assigned into 6 groups of 10 cavities each: (G1) Biodentine, (G2) Fuji II LC as a filling material, (G3) Biodentine as a base + Optibond Solo Plus + silane + Filtek Z250, (G4) as in G3 without silane, (G5) Biodentine as a base + Septobond SE + Filtek Z250, (G6) Fuji II LC as a base + Optibond Solo Plus + Filtek Z250. The materials were applied according to the manufacturers' instructions. Biodentine required no dentin or enamel surface conditioning treatment. The teeth were thermocycled 2500x (5°C to 55°C). The specimens were then sealed with a 1-mm window around the marginal interface. Samples were immersed in a 50% w/v silver nitrate solution and exposed to a photo developing solution. The teeth were embedded in resin (Sody 33) and sectioned through the restorations. The silver penetration was directly measured using a light microscope. The results were expressed as ordinal scores from 0 to 3 at cervical, interfacial, and enamel margins. The data were analyzed with the nonparametric Kruskal-Wallis, Games Howell, and Wilcoxon signed rank tests (p < 0.05). RESULTS: No statistically significant differences were found between the 6 groups, neither for the dentin cervical margins nor for cervical lining (Biodentine or Fuji II LC)/resin composite interfaces. Statistically significant differences were observed between G5 (median score = 2.0) and the other groups (median score = 1.0) for the enamel margins. Statistically significant differences were found between enamel and dentin cervical margins in G2 (enamel median score = 1.0; dentin median score = 1.5) and G5 (enamel median score = 2.0; dentin median score = 1.0). CONCLUSION: Within the limits of this in vitro study, Biodentine as dentin substitute in cervical lining restorations or as a restorative material in approximal cavities when the cervical extent is under the CEJ seems to perform well without any conditioning treatment. However, the operating time is longer than when a RMGIC (Fuji II LC) is used.

Induction of specific cell responses to a Ca(3)SiO(5)-based posterior restorative material.

OBJECTIVES: A Ca(3)SiO(5)-based cement has been developed to circumvent the shortcomings of traditional filling materials. The purpose of this work was to evaluate its genotoxicity, cytotoxicity and effects on the target cells' specific functions. METHODS: Ames' test was applied on four Salmonella typhimurium strains. The micronuclei test was studied on human lymphocytes. The cytotoxicity (MTT test), the Comet assay and the effects on the specific functions by immunohistochemistry were performed on human pulp fibroblasts. RESULTS: Ames' test did not show any evidence of mutagenicity. The incidence of lymphocytes with micronuclei and the percentage of tail DNA in the Comet assay were similar to the negative control. The percentage of cell mortality with the new cement as performed with the MTT test was similar to that of biocompatible materials such as mineral trioxide aggregate (MTA) and was less than that obtained with Dycal. The new material does not affect the target cells' specific functions such as mineralization, as well as expression of collagen I, dentin sialoprotein and Nestin. SIGNIFICANCE: The new cement is biocompatible and does not affect the specific functions of target cells. It can be used safely in the clinic as a single bulk restorative material without any conditioning treatment. It can be used as a potential alternative to traditionally used posterior restorative materials.

Use of two surface analyzers to evaluate the surface roughness of four esthetic restorative materials after polishing.

This study had two aims: determine how well four esthetic restorative materials lent themselves to polishing and compare the results obtained using two different techniques for evaluating surface roughness. The four materials used were two composites modified by the addition of resin, Dyract AP (Dentsply) and Dyract Flow (Dentsply); one composite designed for posterior restorations, SureFil (Dentsply) and one universal micromatrix composite, Esthet-X (Dentsply). Five test pieces were made with each product by inserting the material into cylindrical molds and polymerizing it layer by layer. A single operator polished the specimens on the same day using the Enhance system (Dentsply) and two aluminum oxide pastes. The surfaces were studied successively by means of two surface analyzers: a high-resolution optical profilometer (Nanosurf 488, SAS Technology) and a mechanical profilometer (Mitutoyo Surftest-SV 402). These measurements gave the mean roughness of the surface (Ra). Ten zones were examined for each specimen, and the specimens were observed under an optical microscope (PMG3 inverted metallographic microscope) at 50x magnification. The qualitative and quantitative analyses of the results showed good surface states for all materials. However, the composites based on nano- and micro-filler technology gave the smoothest surfaces after polishing. A comparison of the values obtained with each method of observation showed that mechanical profilometry tended to show roughness caused by polishing, while optical profilometry brought out roughness due to the structure of the material itself.

Activation of human dental pulp progenitor/stem cells in response to odontoblast injury.

In restorative dentistry, whilst moderate carious lesion treatment does not significantly compromise odontoblast cell survival, deep cavity preparation may lead to a partial death of these cells. However, newly formed odontoblast-like cells can replace the necrotic odontoblasts and secrete a reparative dentine matrix. Although several lines of evidence strongly suggest the presence of resting progenitor or stem cells in the dental pulp, little is known about the activation and migration of these cells in response to injury. Human immature third molars extracted for orthodontic reasons were used in this work to study the activation of progenitor/stem cells and their migration after deep cavity preparation involving in pulpal exposure using 5-bromo-2'-deoxyuridine labelling (BrdU). After incubation for 1 day, the BrdU was localised to the nuclei of cells in the perivascular area. The BrdU-immunolabelling exhibited a gradient. It was strong in the blood vessels surrounding the pulpal cavity and decreased in those away from the cavity. After incubation for 2 weeks, labelled cells were seen in the vicinity of the cavity. At 4 weeks, the immunolabelling was localised to the cavity area only. Control teeth without cavities or with shallow dentine cavities did not show any perivascular labelling after culture. These results clearly demonstrate that perivascular progenitor/stem cells can proliferate in response to odontoblast injury. They also show that these proliferating cells can migrate to the pulpal injury site in their tissue of origin simulating the situation in vivo.

Role of injured endothelial cells in the recruitment of human pulp cells.

In restorative dentistry, deep cavity preparation may lead to partial destruction of the odontoblastic layer. However, newly formed odontoblast-like cells can replace the necrotic odontoblasts and secrete a reparative dentine matrix. While growth factors such as transforming growth factor beta1 (TGFbeta1) and bone morphogenetic proteins (BMP-2 and BMP-4) seem to be involved in the proliferation and differentiation of pulp cells, little is known about the migration of the newly proliferating stem cells to the injury site. Our hypothesis was that endothelial cell injury may be involved in directing these cells towards the injury site. For this study, human pulp fibroblasts and L929 cells were fluorescence-labeled by transduction with the Enhanced Green Fluorescent Protein (EGFP). Similarly, human umbilical vein endothelial cells (HUVEC) were labeled with the Discosoma Red Fluorescent Protein-2 (DsRed2). Cell migration was then studied in an insert cell culture system. The HUVEC cells were cultured in the lower compartment while the human pulp fibroblasts or L929 were in the upper compartment. After artificial injury to the HUVEC cells, only human pulp fibroblasts migrated to the lower compartment. At early time periods (4 days), migrating cells were randomly localized on the HUVEC layer. However, after 14 and 20 days, they were perfectly aligned along the injury site. In the absence of injury, no migration was observed. These results suggest that, the endothelial injury is involved in the recruitment of odontoblast-like cells at the injury site.

The biodegradation mechanism of calcium phosphate biomaterials in bone.

This study was undertaken to understand the biodegradation mechanisms of calcium phosphate (Ca-P) biomaterials with different crystallization. Two types of sintered Ca-P porous ceramic (HA and beta-TCP) and a Ca-P bone cement (CPC) were implanted into cavities drilled in rabbit femoral and tibiae condyles. The results have shown that a material biodegradation was rapid in the beta-TCP and the CPC, but very weak in the HA. This biodegradation presented a decrease of material volume from the periphery to the center as well as a particle formation causing phagocytosis by numerous macrophages and multinucleated giant cells in the CPC. In the beta-TCP, there was a peripheral and central decrease of material volume as well as an absence of particle formation or visible phagocytosis. The process of biodegradation is considered to be directly influenced by the type of material crystallization. The sintered bioceramics processed at a high temperature exhibit good crystallization and are primarily degraded by a process dependent on interstitial liquids. However, the bone cement is formed by physicochemical crystallization and is degraded through a dissolution process associated with a cellular process.