acid-phosphatase and Dental-Pulp-Exposure

Interleukin (IL)-17(+) T-helper (Th17) cells and Foxp3(+) regulatory T (Treg) cells are CD4(+) T-helper cells with reciprocal functions in immunology and bone metabolism. The present study aimed to investigate the expression dynamics of Th17 and Treg cells in rat periapical lesions as well as their correlation with bone resorption.. Experimental pulp exposures were made in the lower first molars of 28 Wistar rats to induce periapical lesions. Rats were killed on days 0, 7, 21, and 35. Mandibles were prepared for micro-computed tomography scanning, histologic observation, immunohistochemistry, enzyme histochemistry, and double immunofluorescence analysis.. Through 3-dimensional and 2-dimensional measurements, the volume and area of periapical lesions visibly increased from day 7 to day 21 and then expanded slowly between days 21 and 35. IL-17-positive cells markedly increased from day 7 to day 35. However, Foxp3-positive cells remained at low levels until day 21 and then dramatically increased by day 35. The IL-17(+)/Foxp3(+) ratio and number of osteoclasts simultaneously increased from day 7 to day 21 and then decreased on day 35. Finally, the distinct distribution of CD4(+)/IL-17(+) Th17 and CD4(+)/Foxp3(+) Treg cells was observed on days 7 and 35.. Our findings imply the imbalance of IL-17(+) T cell and Foxp3(+) Treg cell dynamics in induced periapical lesions, which may play an important role in periapical lesion progression.

Topics: Acid Phosphatase; Animals; Bone Resorption; CD4-Positive T-Lymphocytes; Dental Pulp Exposure; Disease Progression; Fluorescent Antibody Technique; Forkhead Transcription Factors; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Interleukin-17; Isoenzymes; Osteoclasts; Periapical Diseases; Rats; Rats, Wistar; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory; Tartrate-Resistant Acid Phosphatase; Th17 Cells; Time Factors; X-Ray Microtomography

Apical periodontitis is an inflammation and destruction of periapical tissues. Matrix metalloproteinase-9 (MMP-9) is thought to be involved in periapical lesion formation and progression. The aim of this study was to evaluate the lesion progression in MMP-9 knockout (KO) mice compared with that in control mice (wild type [WT]).. The pulps of mouse mandibular first molars were exposed; animals were killed at 0, 7, 14, 21, and 28 days after surgery. Hematoxylin-eosin-stained sections were observed for the description of pulpal, apical, periapical features, and the periapical lesion size. The periapical lesion size was further measured with micro-computed tomographic imaging. The number of osteoclasts was also counted by tartrate-resistant acid phosphatase histoenzymology. Real-time polymerase chain reaction and immunohistochemistry were used to analyze the expression levels of receptor activator of NF-κB (RANK), receptor activator of NF-κB ligand (RANKL), osteoprotegerin (OPG), interleukin-1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), MMP-2, and MMP-8.. There was a significant difference (P < .05) between the 2 types of animals regarding the periapical lesion size, which was larger in MMP-9 KO animals. No significant differences (P > .05) were found between WT and MMP-9 KO mice related to the osteoclast number as well as the pulpal, apical, and periapical features. More neutrophil cells were observed in MMP-9 KO animals than WT mice (P < .05). The expression levels of RANK, RANKL, OPG, IL-1β, TNF-α, MMP-2, and MMP-8 were found up-regulated in MMP-9 KO mice (P < .05).. MMP-9 KO animals developed larger periapical lesions with greater inflammatory response, indicating an important role of MMP-9 in the host's immune and inflammatory response to root canal and periradicular infection.

Topics: Acid Phosphatase; Animals; Cell Count; Dental Pulp Exposure; Disease Progression; Interleukin-1beta; Isoenzymes; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Mice; Mice, Knockout; Neutrophils; Osteoclasts; Osteoprotegerin; Periapical Periodontitis; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha; X-Ray Microtomography

The purpose of this study was to evaluate the expression of Dickkopf-1 (DKK-1), a secreted antagonist of the Wnt (wingless)/beta-catenin signaling pathway, during the development of periapical lesions in rats.. Periapical lesions were induced in Wistar rats by occlusal exposure of the pulp of their mandibular first molars. The animals were sacrificed randomly at 0, 7, 14, 21, and 28 day after pulpal exposure. Jaws containing the first molar were obtained and routinely prepared for histologic, immunohistochemical, and enzyme histochemical double immunofluorescence analyses. Data were analyzed using 1-way analysis of variance and Pearson correlation test.. The expansion of the area of periapical lesions was visible from days 7-21 and slowed down thereafter. A few DKK-1- and receptor activator of nuclear factor kappa B ligand (RANKL)-positive cells and osteoclasts were observed on day 7. All positive samples peaked in number on day 14. The expression levels of DKK-1 and RANKL and the number of osteoclasts decreased on days 21 and 28. DKK-1 expression was positively correlated with RANKL expression and osteoclast number from days 7-28.. DKK-1 expression was up-regulated during periapical lesion development. DKK-1 may be associated with the inflammatory response and bone resorption in periapical lesions.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Cell Count; Dental Pulp Exposure; Image Processing, Computer-Assisted; Intercellular Signaling Peptides and Proteins; Isoenzymes; Male; Osteoclasts; Periapical Periodontitis; Periapical Tissue; Random Allocation; RANK Ligand; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Time Factors

Long-term sequential expression of receptor activator of NF-κB ligand (RANKL) and osteoprotegrin (OPG) in lipopolysaccharide (LPS)-induced rat periapical lesions has not been studied..   Seventy-two 4-week-old Wistar rats were divided into eight experimental groups and one control group (eight animals in each).. Lipopolysaccharide-induced periapical lesions were produced in rats by occlusal exposure of the pulp of their lower first molars in all experimental groups but not the control group. The extent of periapical destruction was measured by radiographic imaging. RANKL and OPG mRNA were measured in all tissue sections containing the periapical lesions as well as the control group every week from week 1 to week 8 by real-time quantitative reverse transcription polymerase chain reaction. RANKL and OPG protein were determined by immunohistochemistry. Osteoclasts were identified by enzyme histochemistry.. The sequential changes in the mRNA and protein expression of RANKL and OPG were largely compatible with the occurrence of osteoclasts histologically and enzymes histochemically, as well as the mean areas of the periapical lesions radiographically during long-term observation of the LPS-induced rat periapical lesions.. This study may be the first to demonstrate the long-term RANKL and OPG expression every week from week 1 to week 8 using LPS to produce periapical infection in a Wistar rat model. The long-term findings of high expressions of RANKL and OPG further extend the potential application of the Wistar rat model for future experimental trials using RANKL inhibitor to evaluate the treatment outcome for LPS-induced rat periapical lesions.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Cell Count; Dental Pulp Exposure; Disease Models, Animal; Escherichia coli; Giant Cells; Image Processing, Computer-Assisted; Immunohistochemistry; Isoenzymes; Lipopolysaccharides; Male; Osteoclasts; Osteoprotegerin; Periapical Diseases; Radiography, Bitewing; RANK Ligand; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Tartrate-Resistant Acid Phosphatase; Time Factors

The present study investigated whether bacteria infecting the root canal can activate any infiltrating T cells to produce receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL).. Using a mouse model of periapical lesion induced by artificial dental pulp exposure, the presence of RANKL-positive T cells and osteoclasts in the periapical lesion was examined by an immunohistochemical approach. The bacteria colonizing the exposed root canal were identified by 16S ribosomal RNA (rRNA) sequence analysis. The isolated endodontic bacteria were further immunized to normal mice, and soluble activator of NF-κB ligand (sRANKL) production by the T cells isolated from the immunized mice was evaluated by ex vivo culture system.. RANKL-positive T cells along with TRAP+ osteoclasts were identified in periapical bone resorption lesions. The gram-negative bacterium Pasteurella pnumotropica, which was most frequently detected from the root canal of exposed pulp, showed remarkably elevated serum immunoglobulin G (IgG)-antibody response in pulp-exposed mice compared with control nontreated mice. Immunization of mice with P. pneumotropica induced not only serum IgG-antibody but also primed bacteria-reactive T cells that produced sRANKL in response to ex vivo exposure to P. pneumotropica.. T cells infiltrating the periapical region express RANKL, and the endodontic bacteria colonizing the root canal appear to induce RANKL expression from bacteria-reactive T cells, suggesting the possible pathogenic engagement of the immune response to endodontic bacteria in the context of developing bone resorptive periapical lesions.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Antibodies, Bacterial; Biomarkers; CD3 Complex; Dental Pulp Cavity; Dental Pulp Exposure; Disease Models, Animal; Enterococcus; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Immunization; Immunoglobulin G; Immunologic Memory; Isoenzymes; Male; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Microscopy, Confocal; Osteoclasts; Pasteurella Infections; Pasteurella pneumotropica; Periapical Diseases; RANK Ligand; RNA, Bacterial; RNA, Ribosomal, 16S; T-Lymphocytes; Tartrate-Resistant Acid Phosphatase

The aim of this study was to analyze the contribution of nonresident progenitor/stem cells and hematopoietic cells to reparative dentinogenesis.. Parabiosis was established between C57BL/6-TgN(ACTbEGFP)10sb/J transgenic mice (GFP+) and C57BL/6 wild-type mice (GFP-) to ensure blood cross-circulation between animals. Reparative dentinogenesis was stimulated by pulp exposures and capping on the first maxillary molar in the GFP- mice. Histologic sections of injured molars from GFP- mice were analyzed by epifluorescence microscopy to examine the contributions of GFP+ cells (nonresident progenitor cells and hematopoietic cells originating from GFP+ mice) to reparative dentinogenesis.. GFP+ cells were detected in close association with reparative dentin formed at the site of pulp exposure in the maxillary first molars of the GFP- mice.. The present study suggests the participation of the nonresident progenitor cells and hematopoietic cells in reparative dentinogenesis.

Topics: Acid Phosphatase; Aluminum Compounds; Animals; Biomarkers; Calcium Compounds; Composite Resins; Cross Circulation; Dental Materials; Dental Pulp Capping; Dental Pulp Exposure; Dentin, Secondary; Dentinogenesis; Drug Combinations; Flow Cytometry; Fluorescent Dyes; Green Fluorescent Proteins; Hematopoietic Stem Cells; Isoenzymes; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microscopy, Fluorescence; Models, Animal; Molar; Odontoblasts; Oxides; Parabiosis; Pulp Capping and Pulpectomy Agents; Resin Cements; Silicates; Silicon Dioxide; Stem Cells; Tartrate-Resistant Acid Phosphatase; Zirconium

The purpose of this study was to investigate the activation of p38 mitogen-activated protein kinase (MAPK) during the development of periapical lesions in rats. Periapical lesions developed within 28 days after pulp exposure of mandibular first molars in Wistar rats. The animals were sacrificed randomly at 0, 7, 14, 21, and 28 days after pulpal exposure. The jaws that contained the first molar were obtained and routinely prepared for immunohistochemistry and enzyme histochemistry. A few phosphorylated p38 MAPK (P-p38)-positive cells and osteoclasts could be observed on day 7; both peaked in number on day 14. In the 21- and 28-day samples, the P-p38 MAPK expression decreased and fewer osteoclasts were observed. From day 7 to day 28, a significant positive correlation was found between P-p38 MAPK expression and osteoclasts. These findings showed that the activation of p38 MAPK might be associated with bone resorption in periapical lesions.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Cell Count; Cell Nucleus; Coloring Agents; Dental Pulp Exposure; Enzyme Activation; Immunohistochemistry; Isoenzymes; Male; Mandibular Diseases; Osteoclasts; p38 Mitogen-Activated Protein Kinases; Periapical Diseases; Periapical Periodontitis; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

Nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) plays an important role in host defense, as well as in inflammation-induced tissue lesions. Here we evaluated the role of NO in bone loss in bacterial infection-induced apical periodontitis by using iNOS-deficient mice (iNOS(-/-)). The iNOS(-/-) mice developed greater inflammatory cell recruitment and osteolytic lesions than WT mice. Moreover, tartrate-resistant acid-phosphatase-positive (TRAP(+)) osteoclasts were significantly more numerous in iNOS(-/-) mice. Furthermore, the increased bone resorption in iNOS(-/-) mice also correlated with the increased expression of receptor activator NF-kappaB (RANK), stromal-cell-derived factor-1 alpha (SDF-1 alpha/CXCL12), and reduced expression of osteoprotegerin (OPG). These results show that NO deficiency was associated with an imbalance of bone-resorption-modulating factors, leading to severe infection-stimulated bone loss.

Topics: Acid Phosphatase; Actinomycosis; Alveolar Bone Loss; Animals; Bacterial Infections; Bacteroidaceae Infections; Biomarkers; Cell Count; Cell Movement; Chemokine CXCL12; Dental Pulp Exposure; Isoenzymes; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Nitric Oxide; Nitric Oxide Synthase Type II; Osteoclasts; Osteolysis; Osteoprotegerin; Periapical Periodontitis; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase

The present study was undertaken to determine the frequency and extent of apical root resorption associated with induced periradicular lesions in mice.. Bone and root resorption was quantified by using two- and three-dimensional micro-computed tomography (mu-CT) in the lower first molars of mice subjected to pulp exposure and infection.. mu-CT measurements showed significant apical resorption in exposed and infected teeth, resulting in an average distal root shortening of 12.7% (P <.001 vs unexposed). These findings were confirmed with three-dimensional reconstituted images that showed thinning and shortening of the distal root. Tartrate-resistant acid phosphatase clastic cells were associated with resorption lacunae on the cementum of root apices, as well as on bone at the periphery of the periradicular lesions. Brown and Brenn staining showed the presence of bacteria in dentinal tubules adjacent to resorbed cementum.. Apical root resorption is a prominent and consistent finding associated with periradicular infection in the mouse. This species represents a convenient model for studying the pathogenesis of inflammatory root resorption in vivo.

Topics: Acid Phosphatase; Animals; Biomarkers; Bone Resorption; Coloring Agents; Dental Cementum; Dental Pulp Diseases; Dental Pulp Exposure; Dentin; Disease Models, Animal; Gram-Negative Bacteria; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Isoenzymes; Male; Mandibular Diseases; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Molar; Periapical Diseases; Regression Analysis; Root Resorption; Statistics as Topic; Tartrate-Resistant Acid Phosphatase; Tomography, X-Ray Computed; Tooth Apex; Tooth Root

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Dental Pulp Exposure; Dentin, Secondary; Molar; Rats

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Animals; Dental Pulp; Dental Pulp Exposure; Dogs; Histocytochemistry; NAD; Oxidoreductases; Pyrophosphatases; Succinate Dehydrogenase; Tetrazolium Salts; Thiamine; Wound Healing

Topics: Acid Phosphatase; Albumins; Alkaline Phosphatase; Biological Products; Bone Regeneration; Calcium Hydroxide; Chondroitin; Collagen; Dental Pulp Capping; Dental Pulp Exposure; Dentin, Secondary; Humans; In Vitro Techniques; Odontoblasts; Root Canal Therapy; Sulfates