RESUMO
AIM: To investigate macrophage function in the presence of sustained infection with Enterococcus faecalis, a prevalent root canal resident in asymptomatic apical periodontitis. METHODOLOGY: The human monocyte cell line (THP-1) was differentiated into macrophages by exposure to phorbol myristate acetate (PMA), and the cultures were inoculated with E. faecalis for up to 48 h. At three time-points 90 min, 24 and 48 h after inoculation, the macrophages and their supernatants were examined. Assays included macrophage phagocytosis rate and vitality, bacterial survival, reactive oxygen species (ROS) production, mitochondrial activity, cytokine production and the expression of pro/anti-inflammatory M1/M2 markers. Also, periapical tissue from apicectomy samples of human endodontically treated teeth were collected for histological and immunofluorescent analysis. Statistical differences were analysed with RM ANOVA. RESULTS: E. faecalis were phagocytized, and subsequently, most of the macrophages underwent apoptosis and necrosis. The small population of macrophages that remained vital after 48 h post-inoculation harboured surviving bacteria. Despite a reduction in the number of macrophages over time, the mitochondrial activity of the surviving macrophages remained constant and external ROS decreased, whereas internal ROS increased. During the infection, a shift to a M2 macrophage population at 48 h post-infection was observed; the results were similar to those obtained in periapical human tissue biopsies (p < .05). CONCLUSIONS: The study portrays a continuous non-resolved infection with E. faecalis and activation of macrophages that are polarized to the M2 pro-resolution phenotype.
Assuntos
Enterococcus faecalis , Ativação de Macrófagos , Diferenciação Celular , Humanos , Macrófagos , FagocitoseRESUMO
The use of polytetrafluoroethylene (PTFE) tape as the base layer of temporary restorations had gained popularity mainly due to the ease of manipulation. The aim of this study was to assess whether this method changes the potential for bacterial growth and leakage of temporary restorations. The direct contact test and live/dead fluorescent staining were used for comparing Enterococcus faecalis growth and biofilm formation on PTFE, composite, intermediate restorative material (IRM) and Coltosol F. Confocal laser scanning microscopy was employed to evaluate E. faecalis penetration through 2 mm of PTFE, IRM or Coltosol F placed on the bottom of the pulp chamber and into radicular dentinal tubules in extracted maxillary third molars. The results demonstrated that E. faecalis grows on and penetrates through PTFE significantly more than it does with IRM and Coltosol F, revealing its comparably reduced overall antimicrobial sealing ability when placed as the base part of temporary restorations.
Assuntos
Dente Molar , Politetrafluoretileno , Antibacterianos , Materiais Dentários , Cavidade Pulpar , Enterococcus faecalisRESUMO
BACKGROUND: A devastating condition that leads to trauma-related morbidity, multiple rib fractures, remain a serious unmet clinical need. Systemic administration of mesenchymal stem cells (MSCs) has been shown to regenerate various tissues. We hypothesized that parathyroid hormone (PTH) therapy would enhance MSC homing and differentiation, ultimately leading to bone formation that would bridge rib fractures. METHODS: The combination of human MSCs (hMSCs) and a clinically relevant PTH dose was studied using immunosuppressed rats. Segmental defects were created in animals' fifth and sixth ribs. The rats were divided into four groups: a negative control group, in which animals received vehicle alone; the PTH-only group, in which animals received daily subcutaneous injections of 4 µg/kg teriparatide, a pharmaceutical derivative of PTH; the hMSC-only group, in which each animal received five injections of 2 × 106 hMSCs; and the hMSC + PTH group, in which animals received both treatments. Longitudinal in vivo monitoring of bone formation was performed biweekly using micro-computed tomography (µCT), followed by histological analysis. RESULTS: Fluorescently-dyed hMSCs were counted using confocal microscopy imaging of histological samples harvested 8 weeks after surgery. PTH significantly augmented the number of hMSCs that homed to the fracture site. Immunofluorescence of osteogenic markers, osteocalcin and bone sialoprotein, showed that PTH induced cell differentiation in both exogenously administered cells and resident cells. µCT scans revealed a significant increase in bone volume only in the hMSC + PTH group, beginning by the 4th week after surgery. Eight weeks after surgery, 35% of ribs in the hMSC + PTH group had complete bone bridging, whereas there was complete bridging in only 6.25% of ribs (one rib) in the PTH-only group and in none of the ribs in the other groups. Based on the µCT scans, biomechanical analysis using the micro-finite element method demonstrated that the healed ribs were stiffer than intact ribs in torsion, compression, and bending simulations, as expected when examining bone callus composed of woven bone. CONCLUSIONS: Administration of both hMSCs and PTH worked synergistically in rib fracture healing, suggesting this approach may pave the way to treat multiple rib fractures as well as additional fractures in various anatomical sites.