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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.

Biomarkers↗

[Mineralization of the dental pulp: contributions of tissue engineering to tomorrow's therapeutics in odontology].

When bioactive molecules such as bone sialoprotein (BSP), bone morphogenetic protein-7 (BMP-7, also termed OP-1) and chondrogenic Inducing Agents (CIA, A+4 and A-4) were implanted in the pulp of the first upper molars, mineralizations were induced. They were either limited to the formation of a reparative dentinal bridge closing the pulpal wound (CIA A+4), or filled the mesial part of the coronal pulp (BSP), or filled totally the pulp located in the root canal (BMP-7 and CIA A-4). Consequently, these molecules may change in the next future the every day practice in dentistry.

Animals↗

In vitro characterization of human dental pulp cells: various isolation methods and culturing environments.

Our purpose was to characterize human dental pulp cells isolated by various methods and to examine the behavior of cells grown under various conditions for the purpose of pulp/dentin tissue engineering and regeneration. We compared the growth of human pulp cells isolated by either enzyme digestion or the outgrowth method. Expression of dentin sialophosphoprotein, Cbfa1, and two types of collagen (I and III) in these cells was examined by Western blot or reverse transcription/polymerase chain reaction. Growth of pulp cells on dentin and in collagen gel was also characterized. We found that different isolation methods give rise to different populations or lineages of pulp cells during in vitro passage based on their collagen gene expression patterns. Cells isolated by enzymedigestion had a higher proliferation rate than those isolated by outgrowth. Pulp cells did not proliferate or grew minimally on chemically and mechanically treated dentin surface and appeared to establish an odontoblast-like morphology with a cytoplasmic process extending into a dentinal tubule as revealed by scanning electron microscopy. The contraction of the collagen matrix caused by pulp cells was dramatic: down to 34% on day 14. Our data indicate that (1) the choice of the pulp cell isolation method may affect the distribution of the obtained cell populations, (2) a treated dentin surface might still promote odontoblast differentiation, and (3) a collagen matrix may not be a suitable scaffold for pulp tissue regeneration because of the marked contraction caused by pulp cells in the matrix. The present study thus provides important information and a basis for further investigations pre-requisite to establishing pulp tissue engineering/regeneration protocols.

Base Sequence↗

DNA examination of ancient dental pulp incriminates typhoid fever as a probable cause of the Plague of Athens.

BACKGROUND: Until now, in the absence of direct microbiological evidence, the cause of the Plague of Athens has remained a matter of debate among scientists who have relied exclusively on Thucydides' narrations to introduce several possible diagnoses. A mass burial pit, unearthed in the Kerameikos ancient cemetery of Athens and dated back to the time of the plague outbreak (around 430 BC), has provided the required skeletal material for the investigation of ancient microbial DNA. OBJECTIVE: To determine the probable cause of the Plague of Athens. METHOD: Dental pulp was our material of choice, since it has been proved to be an ideal DNA source of ancient septicemic microorganisms through its good vascularization, durability and natural sterility. RESULTS: Six DNA amplifications targeted at genomic parts of the agents of plague (Yersinia pestis), typhus (Rickettsia prowazekii), anthrax (Bacillus anthracis), tuberculosis (Mycobacterium tuberculosis), cowpox (cowpox virus) and cat-scratch disease (Bartonella henselae) failed to yield any product in 'suicide' reactions of DNA samples isolated from three ancient teeth. On the seventh such attempt, DNA sequences of Salmonella enterica serovar Typhi were identified providing clear evidence for the presence of that microorganism in the dental pulp of teeth recovered from the Kerameikos mass grave. CONCLUSION: The results of this study clearly implicate typhoid fever as a probable cause of the Plague of Athens.

DNA, Bacterial↗

Regulation of interleukin-6 expression in human dental pulp cell cultures stimulated with Prevotella intermedia lipopolysaccharide.

Interleukin (IL)-6 expression in human dental pulp cell cultures after stimulation with Prevotella intermedia lipopolysaccharide (LPS) was investigated by Northern blot analysis, enzyme immunoassay, and bioassay. The IL-6 mRNA expression began to increase after 1 hr and continued after up to 8 hr of exposure on stimulation with 10 microg/ml of P. intermedia LPS. The bioactivity was dose-dependent on the concentration of P. intermedia LPS (0 to 100 microg/ml). The IL-6 mRNA expression was inhibited by actinomysin D and super-induced by cycloheximide. Anti-CD14 monoclonal antibody (MY4) inhibited the IL-6 mRNA expression when administered at a 0.5 microg/ml concentration before stimulation with P. intermedia LPS at 1 microg/ml. The immunoregulatory cytokines (interferon-gamma, IL-10, and IL-4) inhibited LPS-induced IL-6 production with a combined treatment. These results suggest the IL-6 expression by pulp cell cultures is CD14-dependent and regulated at the transcriptional level, and a combined treatment with immunoregulatory cytokines may be effective for control of pulpal inflammation due to P. intermedia LPS.

Antibodies, Monoclonal↗