acid-phosphatase and Periapical-Diseases

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

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

To investigate the effects of systemically administered alendronate, one of the most potent bisphosphonates (BPs), on alveolar bone resorption and angiogenesis in rats subjected to experimental periapical lesions over two time periods.. Forty adult Sprague-Dawley (SD) rats were divided equally into control and experimental groups, and the pulp chambers of mandibular first molars of all rats were exposed to the oral environment to induce periapical lesions. The experimental group received daily subcutaneous injections of alendronate at a dose of 0.25 mg kg(-1), whereas the control group received only the saline vehicle. These injections were initiated 1 week before the periapical lesion induction and then continued daily throughout the entire experimental period. After 2 or 4 weeks following pulp exposure, the rats were killed, and the mandibles were examined histologically for periapical bone loss area, number of microvascular vessels (NMV) and tartrate-resistant acid phosphatase (TRAP) activity.. Overall, periapical bone loss area and the number of TRAP-positive cells (osteoclasts) were significantly decreased at 2 and 4 weeks, respectively, after daily subcutaneous injection of alendronate compared with the control group (P < 0.05). There was no significant decrease change in NMV (P > 0.05).. Administration of alendronate to rats might inhibit alveolar bone resorption associated with periapical disease, which might not lead to impairment of angiogenesis. However, because of the differences between rats and humans, one has to consider the possible consequences of this treatment in the clinic.

Topics: Acid Phosphatase; Alendronate; Alveolar Bone Loss; Alveolar Process; Animals; Bone Density Conservation Agents; Bone Resorption; Disease Models, Animal; Drug Administration Schedule; Isoenzymes; Male; Mandible; Microvessels; Neovascularization, Physiologic; Periapical Diseases; Rats; Rats, Sprague-Dawley; Statistics, Nonparametric; Tartrate-Resistant Acid Phosphatase

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

Lipopolysaccharide (LPS) in the outer layers of Gram-negative bacteria plays an important role in initiating and sustaining periapical lesions. To understand the mechanisms of osteoclastic bone resorption in periapical lesions induced by LPS, we stimulated osteoclast precursors, RAW 264.7 cells with LPS. LPS stimulated osteoclastogenesis when osteoclast precursors were primed with activator for NF-kappaB ligand (RANKL) as little as 24 h. By employing real-time PCR analysis, we have confirmed that osteoclast-like cells stimulated by LPS express high level of osteoclast-specific gene markers such as TRAP, cathepsin K, and calcitonin receptor. These results suggest that bone-resportive action by LPS is partially independent of RANKL.

Topics: Acid Phosphatase; Analysis of Variance; Animals; Biomarkers; Bone Resorption; Carrier Proteins; Cathepsin K; Cathepsins; Cattle; Cell Line; Drug Synergism; Gene Expression; Isoenzymes; Lipopolysaccharides; Membrane Glycoproteins; Mice; Osteoclasts; Periapical Diseases; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Receptors, Calcitonin; Reverse Transcriptase Polymerase Chain Reaction; 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

Attempts were made to detect and measure the activities of arylsulfatases. A&B acid phosphatase, lactate dehydrogenase, and glutamate oxaloacetate transaminase (aspartate transaminase) enzymes in human chronic lesions of endodontic origin. Thirteen periapical lesions of endodontic origin and 11 noninflamed control periapical tissues were obtained. The specimens were carried to the laboratory on liquid nitrogen and kept at -70 degrees C. Samples were thawed, homogenized, and then assayed for enzyme activities. The specific activities of arylsulfatase A (nmol/hr/mg protein) were 55.0+/-10.7 (chronic lesions) vs. 3.4+/-2.2 (controls) (p < 0.01). Arylsulfatase B specific activities (nmol/hr/mg protein) were 50.3+/-6.4 (chronic lesions) vs 91.8+/-18.4 (controls). Total acid phosphatase activities (mU/mg protein) were 45.8+/-6.6 (chronic lesions) vs. 26.8+/-3.1 (controls). Lactate dehydrogenase activities (Berger-Broida units/mg protein) of the chronic periapical lesions were significantly higher than the control group (362+/-63.2) vs. (140+/-46.0) (p < 0.05). There was no significant difference between the specific activities of aspartate transaminase in chronic lesions and the control group (68.0+/-14.5) vs. (53.0+/-10.4) mU/mg protein).

Topics: Acid Phosphatase; Aspartate Aminotransferases; Cerebroside-Sulfatase; Chronic Disease; Dental Pulp Diseases; Humans; L-Lactate Dehydrogenase; N-Acetylgalactosamine-4-Sulfatase; Periapical Diseases; Periapical Tissue; Spectrophotometry; Statistics as Topic