Differential Response of Human Dendritic Cells upon Stimulation with Encapsulated or Non-Encapsulated Isogenic Strains of Porphyromonas gingivalis.

During periodontitis, the extracellular capsule of Porphyromonas gingivalis favors alveolar bone loss by inducing Th1 and Th17 patterns of lymphocyte response in the infected periodontium. Dendritic cells recognize bacterial antigens and present them to T lymphocytes, defining their activation and polarization. Thus, dendritic cells could be involved in the Th1 and Th17 response induced against the P. gingivalis capsule. Herein, monocyte-derived dendritic cells were obtained from healthy individuals and then stimulated with different encapsulated strains of P. gingivalis or two non-encapsulated isogenic mutants. Dendritic cell differentiation and maturation were analyzed by flow cytometry. The mRNA expression levels for distinct Th1-, Th17-, or T-regulatory-related cytokines and transcription factors, as well as TLR2 and TLR4, were assessed by qPCR. In addition, the production of IL-1ß, IL-6, IL-23, and TNF-α was analyzed by ELISA. The encapsulated strains and non-encapsulated mutants of P. gingivalis induced dendritic cell maturation to a similar extent; however, the pattern of dendritic cell response was different. In particular, the encapsulated strains of P. gingivalis induced higher expression of IRF4 and NOTCH2 and production of IL-1ß, IL-6, IL-23, and TNF-α compared with the non-encapsulated mutants, and thus, they showed an increased capacity to trigger Th1 and Th17-type responses in human dendritic cells.

Premature Senescence of T-cells Favors Bone Loss During Osteolytic Diseases. A New Concern in the Osteoimmunology Arena.

Cellular senescence is a biological process triggered in response to time-accumulated DNA damage, which prioritizes cell survival over cell function. Particularly, senescent T lymphocytes can be generated prematurely during chronic inflammatory diseases regardless of chronological aging. These senescent T lymphocytes are characterized by the loss of CD28 expression, a co-stimulatory receptor that mediates antigen presentation and effective T-cell activation. An increased number of premature senescent CD4+CD28- T lymphocytes has been frequently observed in osteolytic diseases, including rheumatoid arthritis, juvenile idiopathic arthritis, ankylosing spondylitis, osteopenia, osteoporosis, and osteomyelitis. Indeed, CD4+CD28- T lymphocytes produce higher levels of osteoclastogenic molecular mediators directly related to pathologic bone loss, such as tumor necrosis factor (TNF)-α, interleukin (IL)-17A, and receptor-activator of nuclear factor κB ligand (RANKL), as compared with regular CD4+CD28+ T lymphocytes. In addition, premature senescent CD8+CD28- T lymphocytes have been negatively associated with bone healing and regeneration by inhibiting osteoblast differentiation and mesenchymal stromal cell survival. Therefore, accumulated evidence supports the role of senescent T lymphocytes in osteoimmunology. Moreover, premature senescence of T-cells seems to be associated with the functional imbalance between the osteolytic T-helper type-17 (Th17) and bone protective T regulatory (Treg) lymphocytes, as well as the phenotypic instability of Treg lymphocytes responsible for its trans-differentiation into RANKL-producing exFoxp3Th17 cells, a key cellular phenomenon directly related to bone loss. Herein, we present a framework for the understanding of the pathogenic characteristics of T lymphocytes with a premature senescent phenotype; and particularly, we revise and discuss their role in the osteoimmunology of osteolytic diseases.

Humanized Mouse Models for the Study of Periodontitis: An Opportunity to Elucidate Unresolved Aspects of Its Immunopathogenesis and Analyze New Immunotherapeutic Strategies.

Periodontitis is an oral inflammatory disease in which the polymicrobial synergy and dysbiosis of the subgingival microbiota trigger a deregulated host immune response, that leads to the breakdown of tooth-supporting tissues and finally tooth loss. Periodontitis is characterized by the increased pathogenic activity of T helper type 17 (Th17) lymphocytes and defective immunoregulation mediated by phenotypically unstable T regulatory (Treg), lymphocytes, incapable of resolving the bone-resorbing inflammatory milieu. In this context, the complexity of the immune response orchestrated against the microbial challenge during periodontitis has made the study of its pathogenesis and therapy difficult and limited. Indeed, the ethical limitations that accompany human studies can lead to an insufficient etiopathogenic understanding of the disease and consequently, biased treatment decision-making. Alternatively, animal models allow us to manage these difficulties and give us the opportunity to partially emulate the etiopathogenesis of periodontitis by inoculating periodontopathogenic bacteria or by placing bacteria-accumulating ligatures around the teeth; however, these models still have limited translational application in humans. Accordingly, humanized animal models are able to emulate human-like complex networks of immune responses by engrafting human cells or tissues into specific strains of immunodeficient mice. Their characteristics enable a viable time window for the study of the establishment of a specific human immune response pattern in an in vivo setting and could be exploited for a wider study of the etiopathogenesis and/or treatment of periodontitis. For instance, the antigen-specific response of human dendritic cells against the periodontopathogen Porphyromonas gingivalis favoring the Th17/Treg response has already been tested in humanized mice models. Hypothetically, the proper emulation of periodontal dysbiosis in a humanized animal could give insights into the subtle molecular characteristics of a human-like local and systemic immune response during periodontitis and support the design of novel immunotherapeutic strategies. Therefore, the aims of this review are: To elucidate how the microbiota-elicited immunopathogenesis of periodontitis can be potentially emulated in humanized mouse models, to highlight their advantages and limitations in comparison with the already available experimental periodontitis non-humanized animal models, and to discuss the potential translational application of using these models for periodontitis immunotherapeutics.

Extrusión Apical de Barro Dentinario e Irrigante Producidos por Dos Sistemas de Instrumentación de Níquel Titanio al Utilizar Irrigación Pasiva o Activa / Apical Extrusion of Dentin and Irrigant Smear Layer Produced by Two Nickel Titanium Instrumentation Systems when Using Passive or Active Irrigation

RESUMEN: El barro dentinario producido durante tratamiento endodóntico puede ser extruido hacia el tejido periradicular junto con el irrigante, produciendo inflamación y dolor postoperatorio. Comparar la cantidad de extrusión apical de barro dentinario e irrigante, producido durante la preparación químico-mecánica, por dos sistemas de instrumentación Rotatoria (Mtwo), y Reciprocante (Reciproc), complementado con irrigación pasiva o activa (Endoactivator). Cuarenta y ocho premolares (48) inferiores, fueron aleatoriamente distribuidos en cuatro grupos de estudio (n=12 dientes), (1) Rotatorio-pasiva, (2) Reciprocante-pasiva, (3) Rotatorio-activa y (4) Reciprocante-activa. Los conductos radiculares fueron instrumentados de acuerdo a las instrucciones del fabricante, e irrigados con hipoclorito de sodio al 5 %. El barro dentinario e irrigante extruido, fueron recolectados en tubos Eppendorf previamente pesados. Se calcularon los valores de extrusión de barro dentinario e irrigante para cada grupo. Los datos fueron analizados con el análisis de varianza ANOVA. En todos los grupos se produjo extrusión apical. No se encontraron diferencias estadísticamente significativas en la extrusión apical de barro dentinario, entre los grupos estudiados (P = 0,068), sin embargo, al analizar la extrusión de irrigantes, se evidenció diferencia estadísticamente significativa entre los grupos 1 y 4 (P< 0,05), entre los demás grupos no se observaron diferencias estadísticamente significativas. (P > 0,05). Bajo las condiciones de este estudio, los sistemas rotarios y reciprocante, extruyen sólido y líquido, siendo el sistema reciprocante el que produce mayor extrusión de líquido, tanto con irrigación activa como pasiva.

ABSTRACT: The smear layer produced during endodontic treatment can be extruded into the periradicular tissue together with the irrigant, producing inflammation and postoperative pain. The purpose of this in vitro study was to compare the amount of apical extrusion, produced during the endodontic preparations with two rotary instrumentation systems using passive and active irrigation. Forty-eight (48) mandibular premolars were randomly assigned to 4 groups (n = 12 teeth), (1) Rotatory - passive, (2) Reciprocating passive, (3) Rotatory-active, (4) Reciprocating- active. The root canals were instrumented according to the manufacturer's instructions and irrigated with 5 % sodium hypochlorite. Extruded debris and irrigant were collected in previously weighed Eppendorf tubes. The extrusion values were calculated for each group. The data were analyzed with the ANOVA analysis of variance. Results: Apical extrusion was produced in all groups. No statistically significant differences were found in the apical extrusion of smear layer, between the groups studied (P = 0.068), however, when analyzing the extrusion of irrigants, a statistically significant difference was evidenced between groups 1 and 4 (P <0.05). , among the other groups no statistically significant differences were observed. (P> 0.05). Under the conditions of this study, the reciprocating and rotary systems, extrude solid and liquid, being the reciprocating system the one that produces more extrusion of liquid, with both active and passive irrigation.