ABSTRACT
A large number of experimental studies has demonstrated that angiotensin II (Ang II) is involved in key events of the inflammatory process. This study aimed to evaluate the role of Ang II type 1 (AT1) and Ang II type 2 (AT2) receptors on periodontitis. Methods: Experimental periodontitis was induced by placing a 5.0 nylon thread ligature around the second upper left molar of AT1 mice, no-ligature or ligature (AT1-NL and AT1-L), AT2 (AT2-NL or AT2-L) and wild type (WT-NL or L). Alveolar bone loss was scanned using Micro-CT. Cytokines, peptides and enzymes were analyzed from gingival tissues by Elisa and RT-PCR. Results: The blockade of AT1 receptor resulted in bone loss, even in healthy animals. Ang II receptor blockades did not prevent linear bone loss. Ang II and Ang 1-7 levels were significantly increased in the AT2-L (p < 0.01) group compared to AT2-NL and AT1-L. The genic expression of the Mas receptor was significantly increased in WT-L and AT2-L compared to (WT-NL and AT2-NL, respectively) and in AT1-L. Conclusions: Our data suggest that the receptor AT1 appears to be important for the maintenance of bone mass. AT2 receptor molecular function in periodontitis appears to be regulated by AT1.
Subject(s)
Alveolar Bone Loss/metabolism , Mandibular Diseases/metabolism , Periodontitis/metabolism , Receptor, Angiotensin, Type 1/metabolism , Receptor, Angiotensin, Type 2/metabolism , Alveolar Bone Loss/genetics , Alveolar Bone Loss/pathology , Angiotensin II/metabolism , Animals , Disease Models, Animal , Male , Mandibular Diseases/genetics , Mandibular Diseases/pathology , Mice , Mice, Knockout , Periodontitis/genetics , Periodontitis/pathology , Receptor, Angiotensin, Type 1/genetics , Receptor, Angiotensin, Type 2/geneticsABSTRACT
Mandibular osteomyelitis (OM) is a challenging disease. Our objective was to assess a new OM model in rabbits induced by arsenic trioxide and to assess the efficacy of local treatment of OM using injectable gentamicin-collagen hydrogels (GNT-COLL). OM was induced unilaterally by controlled confinement of arsenic trioxide paste to the root canal of lower incisors of rabbits, while OM progression was characterized for 16 weeks. On the other hand, two injectable COLL hydrogels functionalized with GNT were prepared and characterized for physicochemical properties; a simple GNT-COLL and a nanohydroxyapatite (nHA)- loaded hydrogel (GNT-COLL/nHA). The two hydrogels were evaluated to treat OM model, while a multidose intramuscular GNT solution served as positive control. Outcomes were assessed by standard methods at 4 and 12 weeks post-surgery. The clinical, radiographical, and histopathological findings provided evidence for the validity of the arsenic-induced OM. The results demonstrated that a single intra-lesional injection of the two hydrogels was more suppressive to OM compared to multidose systemic GNT. The composite GNT-COLL/nHA hydrogel proved to induce early preservation of alveolar bone (ridge) length and higher amount of bone area\total area at 4 weeks (40.53% ± 2.34) followed by GNT-COLL (32.21% ± 0.72). On the other hand, the positive control group revealed the least ridge length and bone area\total area (26.22% ± 1.32) at 4 weeks. Both hydrogels successfully arrested OM with no signs of recurrence for up to 12 weeks. Therefore, results support the greater advantages of the composite hydrogel as an osteogenic/antibiotic delivery system in OM treatment.
Subject(s)
Biomimetic Materials/pharmacology , Drug Carriers/pharmacology , Gentamicins/pharmacology , Hydrogels/pharmacology , Mandibular Diseases/drug therapy , Osteomyelitis/drug therapy , Animals , Biomimetic Materials/chemistry , Disease Models, Animal , Drug Carriers/chemistry , Gentamicins/chemistry , Hydrogels/chemistry , Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Osteomyelitis/metabolism , Osteomyelitis/pathology , RabbitsABSTRACT
OBJECTIVES: The aim of this study was to investigate the role of p75 neurotrophin receptor (p75NTR) in regulating the mouse alveolar bone development and the mineralization potential of murine ectomesenchymal stem cells (EMSCs). Moreover, we tried to explore the underlying mechanisms associated with the PI3K/Akt/ß-catenin pathway. MATERIALS AND METHODS: p75NTR knockout (p75NTR-/- ) mice and wild-type (WT) littermates were used. E12.5d p75NTR-/- and WT EMSCs were isolated in the same pregnant p75NTR-/+ mice from embryonic maxillofacial processes separately. Mouse alveolar bone mass was evaluated using micro-CT. Differential osteogenic differentiation pathways between p75NTR-/- and WT EMSCs were analysed by RNA-sequencing. The PI3K inhibitor LY294002 and PI3K agonist 740Y-P were used to regulate the PI3K/Akt pathway in EMSCs. p75NTR overexpression lentiviruses, p75NTR knock-down lentiviruses and recombined mouse NGF were used to transfect cells. RESULTS: The alveolar bone mass was found reduced in the p75NTR knockout mouse comparing to the WT mouse. During mineralization induction, p75NTR-/- EMSCs displayed decreased osteogenic capacity and downregulated PI3K/Akt/ß-catenin signalling. The PI3K/Akt/ß-catenin pathway positively regulates the potential of differential mineralization in EMSCs. The promotive effect of p75NTR overexpression can be attenuated by LY294002, while the inhibitory effect of p75NTR knock-down on Runx2 and Col1 expression can be reversed by 740Y-P. CONCLUSION: Deletion of p75NTR reduced alveolar bone mass in mice. P75NTR positively regulated the osteogenic differentiation of EMSCs via enhancing the PI3K/Akt/ß-catenin pathway.
Subject(s)
Alveolar Bone Loss/genetics , Mandibular Diseases/genetics , Receptors, Nerve Growth Factor/genetics , Signal Transduction , Alveolar Bone Loss/metabolism , Alveolar Bone Loss/pathology , Animals , Cells, Cultured , Gene Deletion , Male , Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Mice , Mice, Knockout , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Receptors, Nerve Growth Factor/metabolism , beta Catenin/metabolismABSTRACT
The importance of considering bone quality during oral implant treatment is increasingly being recognized. Assessment of bone quality in response to changes in the jaw bone is extremely important when planning treatment. The present study analyzed biological apatite (BAp) crystallites, a bone quality factor, in order to investigate crystallographic anisotropy in dentate and edentulous human mandibles. Using mandibular samples from Japanese adult cadavers, a region of interest was established comprising cortical bone in the central incisors. Samples were classified into five morphological categories based on the extent of bone resorption. Bone mineral density (BMD) was measured and diffraction intensity ratios were calculated using a microbeam X-ray diffraction system. While no differences were observed in BMD, differences were observed in BAp crystallite alignment between the measurement points. In the alveolar region, samples with residual alveolar bone showed strong alignment in the occlusal direction, while samples with marked alveolar bone resorption had preferential alignment in the mesiodistal direction. The present findings suggest that tooth loss and the extent of alveolar bone resorption affects bone quality in the mandible. © 2018 Wiley Periodicals, Inc. J. Biomed. Mater. Res. Part B: 2018. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 838-846, 2019.
Subject(s)
Alveolar Bone Loss , Apatites/metabolism , Bone Density , Dental Implants , Mandible , Mandibular Diseases , Aged , Aged, 80 and over , Alveolar Bone Loss/diagnostic imaging , Alveolar Bone Loss/metabolism , Female , Humans , Male , Mandible/diagnostic imaging , Mandible/metabolism , Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Middle AgedABSTRACT
The purpose of the study was to perform an immunohistochemical and histological evaluation of samples taken from different bone regeneration procedures in atrophic human mandible. 30 patients (15 men and 15 women, age range of 35-60 years), non-smokers, with good general and oral health were recruited in this study and divided into three groups. The first group included patients who were treated with blood Concentration Growth Factors (bCGF), the second group included patients who were treated with a mixture of bCGF and autologous bone, while the third group of patients was treated with bCGF and tricalcium phosphate/hydroxyapatite (TCP-HA). Six months after the regenerative procedures, all patients undergone implant surgery, and a bone biopsy was carried out in the site of implant insertion. Each sample was histologically and immunohistochemically examined. Histological evaluation showed a complete bone formation for group II, partial ossification for group I, and moderate ossification for group III. Immunohistochemical analysis demonstrated a statistically significant difference between the three groups, and the best clinical result was obtained with a mixture of bCGF and autologous bone.
Subject(s)
Bone Remodeling/drug effects , Bone Transplantation , Hydroxyapatites/therapeutic use , Immunohistochemistry , Intercellular Signaling Peptides and Proteins/therapeutic use , Mandible/drug effects , Mandible/surgery , Mandibular Diseases/therapy , Adult , Atrophy , Biopsy , Dental Implantation , Europe , Female , Humans , Male , Mandible/metabolism , Mandible/pathology , Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Middle Aged , Prospective Studies , Time Factors , Transplantation, Autologous , Treatment OutcomeABSTRACT
Jaw discrepancies and malrelations affect a large proportion of the general population and their treatment is of utmost significance for individuals' health and quality of life. The aim of their therapy is the modification of aberrant jaw development mainly by targeting the growth potential of the mandibular condyle through its cartilage, and the architectural shape of alveolar bone through a suture type of structure, the periodontal ligament. This targeted treatment is achieved via external mechanical force application by using a wide variety of intraoral and extraoral appliances. Condylar cartilage and sutures exhibit a remarkable plasticity due to the mechano-responsiveness of the chondrocytes and the multipotent mesenchymal cells of the sutures. The tissues respond biologically and adapt to mechanical force application by a variety of signaling pathways and a final interplay between the proliferative activity and the differentiation status of the cells involved. These targeted therapeutic functional alterations within temporo-mandibular joint ultimately result in the enhancement or restriction of mandibular growth, while within the periodontal ligament lead to bone remodeling and change of its architectural structure. Depending on the form of the malrelation presented, the above treatment approaches, in conjunction or separately, lead to the total correction of jaw discrepancies and the achievement of facial harmony and function. Overall, the treatment of craniofacial and jaw anomalies can be seen as an interplay of mechanical forces and adaptations occurring within temporo-mandibular joint and alveolar bone. The aim of the present review is to present up-to-date knowledge on the mechano-biology behind jaw growth modification and alveolar bone remodeling. Furthermore, future molecular targeted therapeutic strategies are discussed aiming at the improvement of mechanically-driven chondrogenesis and osteogenesis.
Subject(s)
Mandible , Mandibular Diseases , Stress, Mechanical , Animals , Bone Remodeling , Cell Proliferation , Humans , Mandible/metabolism , Mandible/pathology , Mandible/physiopathology , Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Mandibular Diseases/physiopathology , Mandibular Diseases/therapy , Periodontal Ligament/metabolism , Periodontal Ligament/pathology , Periodontal Ligament/physiopathology , Temporomandibular Joint/metabolism , Temporomandibular Joint/pathology , Temporomandibular Joint/physiopathologyABSTRACT
A tissue engineering approach to address craniofacial defects requires a biomaterial that balances macro-scale mechanical stiffness and strength with the micron-scale features that promote cell expansion and tissue biosynthesis. Such criteria are often in opposition, leading to suboptimal mechanical competence or bioactivity. We report the use of a multiscale composite biomaterial that integrates a polycaprolactone (PCL) reinforcement structure with a mineralized collagen-glycosaminoglycan scaffold to circumvent conventional tradeoffs between mechanics and bioactivity. The composite promotes activation of the canonical bone morphogenetic protein 2 (BMP-2) pathway and subsequent mineralization of adipose-derived stem cells in the absence of supplemental BMP-2 or osteogenic media. We subsequently examined new bone infill in the acellular composite, scaffold alone, or PCL support in 10 mm dia. ramus mandibular defects in Yorkshire pigs. We report an analytical approach to quantify radial, angular, and depth bone infill from micro-computed tomography data. The collagen-PCL composite showed improved overall infill, and significantly increased radial and angular bone infill versus the PCL cage alone. Bone infill was further enhanced in the composite for defects that penetrated the medullary cavity, suggesting recruitment of marrow-derived cells. These results indicate a multiscale mineralized collagen-PCL composite offers strategic advantages for regenerative repair of craniofacial bone defects.
Subject(s)
Collagen/chemistry , Mandibular Diseases/drug therapy , Polyesters/chemistry , Animals , Bone and Bones/pathology , Mandibular Diseases/metabolism , Swine , Wound Healing/drug effectsABSTRACT
Skeletal mandibular hypoplasia (SMH), one of the common types of craniofacial deformities, seriously affects appearance, chewing, pronunciation, and breathing. Moreover, SMH is prone to inducing obstructive sleep apnea syndrome. We found that brain and muscle ARNT-like 1 (BMAL1), the core component of the molecular circadian oscillator, was significantly decreased in mandibles of juvenile SMH patients. Accordingly, SMH was observed in circadian-rhythm-disrupted or BMAL1-deficient mice. RNA sequencing and protein chip analyses suggested that matrix metallopeptidase 3 (MMP3) is the potential target of BMAL1. Interestingly, in juvenile SMH patients, we observed that MMP3 was obviously increased. Consistently, MMP3 was upregulated during the whole growth period of 3-10 weeks in Bmal1-/- mice. Given these findings, we set out to characterize the underlying mechanism and found BMAL1 deficiency enhanced Mmp3 transcription through activating p65 phosphorylation. Together, our results provide insight into the mechanism by which BMAL1 is implicated in the pathogenesis of SMH.
Subject(s)
ARNTL Transcription Factors/deficiency , Gene Expression Regulation, Enzymologic , Mandibular Diseases/metabolism , Matrix Metalloproteinase 3/biosynthesis , Up-Regulation , Animals , Child , Female , Humans , Male , Mandibular Diseases/genetics , Mandibular Diseases/pathology , Matrix Metalloproteinase 3/genetics , Mice , Mice, KnockoutABSTRACT
BACKGROUND: Focal adhesion kinase (FAK) is an important mediator of cell adhesion, growth proliferation, survival, angiogenesis, and migration. FAK is overexpressed in many locally invasive and malignant lesions including oral cancer. Looking at the tumorigenic nature of keratocystic odontogenic tumor (KCOT), which involves local invasion, proliferation, and recurrence, we hypothesized strong expression of FAK in the epithelial lining of KCOT. MATERIALS AND METHODS: 34 KCOTs, 11 orthokeratinized odontogenic cysts (OOCs), 25 radicular cysts (RCs), 17 dentigerous cysts (DCs), and 25 dental follicles (DFs) were retrieved from archives and subjected to the immunohistochemical analysis using FAK antibody. RESULTS: In KCOT, strong expression was observed in 22 (62.8%) cases followed by weak and negative expression in 9 (25.71%) and 4 (11.4%) cases, respectively. Negative expression was seen in 7 (63.63%) cases of OOC, while 4 (36.36%) showed weak expression. In case of RC, 20 (80%) cases displayed negative expression and 4 (16%) and 1 (4%) cases showed weak and strong expressions, respectively. In case of DC, negative expression was seen in 14 (82.35%) cases and weak expression in 3 (17.64%) cases. DF was characterized by negative [21 (84%)] and weak expression [4 (16%)]. Nuclear expression of FAK was seen only in KCOT (11 cases). There was statistically significant higher FAK expression in KCOT as compared to OOC, RC, DC, and DF (P < 0.00001). CONCLUSION: FAK molecule could be an important player in tumorigenesis of KCOT and thus is a potential target for future drug development.
Subject(s)
Focal Adhesion Protein-Tyrosine Kinases/metabolism , Mandibular Diseases/metabolism , Maxillary Diseases/metabolism , Odontogenic Cysts/metabolism , Adolescent , Adult , Child , Female , Humans , Male , Young AdultABSTRACT
Glandular odontogenic cyst (GOC) is a relatively rare but well-described clinicopathologic entity. Its rarity and unpredictable clinical behavior are challenging to managing clinicians. Its variable and overlapping histomorphologic features are also diagnostically challenging for pathologists. Other odontogenic cysts and oral cystic neoplasms can simulate GOC. There are specific histologic criteria that help distinguish GOC from other mimickers. To our knowledge, the phenomenon of hemosiderin pigments deposition within the lining glandular epithelium of GOC has not been covered in detail or specifically reported so far in the literature. We report a case of nontraumatized anterior mandibular GOC in a middle-aged male, which histologically showed hemosiderin pigments within the lining epithelium without stromal siderophages. This finding might reflect a nonspecific spontaneous intraluminal hemorrhage. However, intraepithelial hemosiderin in GOC may be an additional helpful diagnostic clue of GOC in challenging cases since this phenomenon has not been reported in other mimicker cystic lesions.
Subject(s)
Hemosiderin/metabolism , Mandibular Diseases/pathology , Odontogenic Cysts/pathology , Adult , Epithelium/metabolism , Epithelium/pathology , Humans , Male , Mandibular Diseases/metabolism , Middle Aged , Odontogenic Cysts/metabolismABSTRACT
The long-term stability and qualitative characteristics of periodontium regenerated by FGF-2 treatment were compared with normal physiological healing tissue controls in a Beagle dog 2-wall periodontal defect model 13 months after treatment by assessing tissue histology and three-dimensional microstructure using micro-computed tomography (µCT). After FGF-2 (0.3%) or vehicle treatment at the defect sites, serial changes in the bone mineral content (BMC) were observed using periodic X-ray imaging. Tissues were harvested at 13 months, evaluated histomorphometrically, and the cortical bone volume and trabecular bone structure of the newly formed bone were analyzed using µCT. FGF-2 significantly increased the BMC of the defect area at 2 months compared with that of the control group, and this difference was unchanged through 13 months. The cortical bone volume was significantly increased by FGF-2, but there was no difference between the groups in trabecular bone structure. Bone maturation was occurring in both groups because of the lower cortical volume and denser trabecular bone than what is found in intact bone. FGF-2 also increased the area of newly formed bone as assessed histomorphometrically, but the ratios of trabecular bone in the defect area were similar between the control and FGF-2 groups. These results suggest that FGF-2 stimulates neogenesis of alveolar bone that is of similar quality to that of the control group. The lengths of the regenerated periodontal ligament and cementum, measured as the distance from the defect bottom to the apical end of the gingival epithelium, and height and area of the newly formed bone in the FGF-2 group were larger than those in the control group. The present study demonstrated that, within the limitation of artificial periodontal defect model, the periodontal tissue regenerated by FGF-2 was maintained for 13 months after treatment and was qualitatively equivalent to that generated through the physiological healing process.
Subject(s)
Fibroblast Growth Factor 2/pharmacology , Mandibular Diseases/metabolism , Periodontal Ligament/drug effects , Periodontium/drug effects , Periodontium/pathology , Animals , Bone Density/physiology , Bone and Bones/drug effects , Bone and Bones/pathology , Dogs , Female , Mandibular Diseases/etiologyABSTRACT
BACKGROUND: Central giant-cell lesions (CGCLs) are reactive lesions that consist histologically of spindle-shaped stromal cells, (fibroblasts and myofibroblasts) loosely arranged in a fibrous stroma, multinucleated giant cells and mononuclear cells with haemorrhagic areas. This study identified the immunoexpression of alpha-smooth muscle actin in spindle-shaped stromal cells, and glucocorticoid and calcitonin receptors in multinucleated giant cells and mononuclear cells. Their association with the clinical and radiographic characteristics of these lesions was identified. METHODS: Thirty-five cases of CGCLs were studied. Expression of alpha-smooth muscle actin, glucocorticoid and calcitonin was evaluated by immunohistochemistry. The labelling index was 100 times the quotient of the number of positive cells divided by the total number of cells of each type. Logistic regression analysis was applied. RESULTS: Alpha-smooth muscle actin was positive (54%) for spindle stromal cells (myofibroblasts). A significant association was observed with root resorption (P = 0.004) and cortical bone destruction (P = 0.024). Glucocorticoid immunoexpression was positive for 99% of the giant cells and 86.7% of the mononuclear cells. Glucocorticoid immunoexpression in the mononuclear cells was associated with root resorption (P = 0.031). A longer evolution time was associated with lower immunoexpression of glucocorticoid (OR 12.4: P = 0.047). Calcitonin immunoexpression was positive in 86% of the giant cells. Immunoexpression of calcitonin was associated with age (P = 0.040). CONCLUSIONS: Myofibroblasts are important components of CGCLs, stromal cells and alpha-smooth muscle. Actin immunoexpression was associated with root and cortical bone resorption.
Subject(s)
Actins/biosynthesis , Granuloma, Giant Cell/metabolism , Mandibular Diseases/metabolism , Maxillary Diseases/metabolism , Receptors, Calcitonin/biosynthesis , Receptors, Glucocorticoid/biosynthesis , Adolescent , Adult , Aged , Aged, 80 and over , Bone Resorption/pathology , Child , Cross-Sectional Studies , Female , Granuloma, Giant Cell/pathology , Humans , Immunohistochemistry , Male , Mandibular Diseases/pathology , Maxillary Diseases/pathology , Middle Aged , Myofibroblasts/metabolism , Myofibroblasts/pathology , Stromal Cells/metabolism , Stromal Cells/pathology , Young AdultABSTRACT
PURPOSE: To determine the biological behaviour of common odontogenic cystic lesions by analysing and comparing bcl-2 expression amongst them. MATERIALS AND METHODS: Our study covered 90 formalin fixed paraffin embedded tissue samples: 26 primary cases each of radicular cysts (RC), dentigerous cysts (DC) and odontogenic keratocysts (OKC) and 12 of recurrent OKCs. Bcl-2 expression was analysed immunohistochemically and data analysis was accomplished using SPSS version 17.0. Means were taken for age while for gender and site of the lesions frequencies and percentages were determined. The Chi-square test was applied to evaluate any statistically significant difference of bcl-2 expression in these lesions and p value of ≤0.05 was taken as significant. RESULTS: All the recurrent OKCs showed a strong positivity for bcl-2 that was absent in all of its primary cases (p value<0.05). Although variation in expression of bcl-2 was not found to be statistically significant between RC and DC, however, it became significant when all primary cases of these common odontogenic lesions were compared. CONCLUSIONS: Recurrent OKC showed comparatively a more aggressive behaviour than their primary counterparts and also from RC and DC. Bcl-2 proved to be a valuable adjunct in determining aggressive biological behaviour of odontogenic lesions.
Subject(s)
Mandibular Diseases/metabolism , Maxillary Diseases/metabolism , Odontogenic Cysts/chemistry , Proto-Oncogene Proteins c-bcl-2/analysis , Adolescent , Adult , Aged , Child , Child, Preschool , Dentigerous Cyst/chemistry , Female , Humans , Immunohistochemistry , Male , Middle Aged , Radicular Cyst/chemistry , Recurrence , Young AdultABSTRACT
Periodontal disease (Periodontitis) is a serious disease that affects a majority of adult Americans and is associated with other systemic diseases, including diabetes, rheumatoid arthritis, and other inflammatory diseases. While great efforts have been devoted toward understanding the pathogenesis of periodontitis, there remains a pressing need for developing potent therapeutic strategies for targeting this pervasive and destructive disease. In this study, we utilized novel adeno-associated virus (AAV)-mediated Atp6v1c1 knockdown gene therapy to treat bone erosion and inflammatory caused by periodontitis in mouse model. Atp6v1c1 is a subunit of the V-ATPase complex and regulator of the assembly of the V0 and V1 domains of the V-ATPase complex. We demonstrated previously that Atp6v1c1 has an essential function in osteoclast mediated bone resorption. We hypothesized that Atp6v1c1 may be an ideal target to prevent the bone erosion and inflammation caused by periodontitis. To test the hypothesis, we employed AAV RNAi knockdown of Atp6v1c1 gene expression to prevent bone erosion and gingival inflammation simultaneously. We found that lesion-specific injection of AAV-shRNA-Atp6v1c1 into the periodontal disease lesions protected against bone erosion (>85%) and gingival inflammation caused by P. gingivalis W50 infection. AAV-mediated Atp6v1c1 knockdown dramatically reduced osteoclast numbers and inhibited the infiltration of dendritic cells and macrophages in the bacteria-induced inflammatory lesions in periodontitis. Silencing of Atp6v1c1 expression also prevented the expressions of osteoclast-related genes and pro-inflammatory cytokine genes. Our data suggests that AAV-shRNA-Atp6v1c1 treatment can significantly attenuate the bone erosion and inflammation caused by periodontitis, indicating the dual function of AAV-shRNA-Atp6v1c1 as an inhibitor of bone erosion mediated by osteoclasts, and as an inhibitor of inflammation through down-regulation of pro-inflammatory cytokine expression. This study demonstrated that Atp6v1c1 RNAi knockdown gene therapy mediated by AAV-shRNA-Atp6v1c1 is a promising novel therapeutic approach for the treatment of bone erosion and inflammatory related diseases, such as periodontitis and rheumatoid arthritis.
Subject(s)
Alveolar Bone Loss/genetics , Inflammation/genetics , Mandibular Diseases/genetics , Periodontitis/genetics , Vacuolar Proton-Translocating ATPases/genetics , Alveolar Bone Loss/immunology , Alveolar Bone Loss/metabolism , Alveolar Bone Loss/therapy , Animals , Disease Models, Animal , Female , Genetic Therapy , Inflammation/immunology , Inflammation/metabolism , Inflammation/therapy , Mandibular Diseases/immunology , Mandibular Diseases/metabolism , Mandibular Diseases/therapy , Mice , Mice, Inbred BALB C , Mice, Knockout , Osteoclasts/drug effects , Osteoclasts/physiology , Periodontitis/immunology , Periodontitis/metabolism , Periodontitis/therapy , RNA, Small Interfering/genetics , RNA, Small Interfering/pharmacology , Vacuolar Proton-Translocating ATPases/antagonists & inhibitorsABSTRACT
BACKGROUND AND OBJECTIVE: Modeling of periodontal bone regeneration in a large animal enables better examination of the spatial and temporal regulation of osteogenesis and the remodeling of the healing defect. RANK, RANKL and osteoprotegerin (OPG) are known to be important regulators of bone healing. The aim of this study was to create periodontal defects surgically in a large animal model and to examine bone regeneration and the expression of RANK, RANKL and OPG proteins in the defect site during bone regeneration. MATERIAL AND METHODS: Periodontal defects were made in the furcation of the second mandibular premolar of sheep. Wound healing was examined 6 h, and 1, 4 and 6 wk after surgery and in control tissue. The teeth and defect region were decalcified and paraffin embedded. Immunohistochemistry for RANK, RANKL and OPG was conducted. Osteoclasts were identified using TRAP staining. RESULTS: The defects were examined at different time points after surgery and by 6 wk the defect region had fully regenerated with new bone, albeit less dense than that in the unwounded controls. RANK-positive osteoclasts were present at the edge of the wound from week 1 and were found within the defect at week 6, corresponding to osteoclast activation and bone remodeling. RANKL staining increased from week 1 compared with unwounded tissue, and peaked at 4 and 6 wk, as the osteoblast numbers increased. At the same time, OPG immunostaining was high in controls and at week 6, suggesting that it may act to block RANKL and control the bone remodeling within the defect. CONCLUSION: Distinctive temporal and spatial expression patterns for RANK, RANKL and OPG proteins were observed during healing of surgically created periodontal wounds in a sheep model. The research identifies possible therapeutic approaches to periodontal bone repair via modulation of these members of the tumor necrosis factor family.
Subject(s)
Furcation Defects/metabolism , Osteoprotegerin/analysis , RANK Ligand/analysis , Receptor Activator of Nuclear Factor-kappa B/analysis , Animals , Bicuspid/pathology , Bone Density/physiology , Bone Regeneration/physiology , Bone Remodeling/physiology , Connective Tissue/pathology , Disease Models, Animal , Female , Furcation Defects/pathology , Mandibular Diseases/metabolism , Osteoblasts/pathology , Osteoclasts/pathology , Osteogenesis/physiology , Sheep , Tartrate-Resistant Acid Phosphatase/analysis , Time Factors , Wound Healing/physiologyABSTRACT
PURPOSE OF REVIEW: Bone disease is a leading cause of fractures and continues to be a source of significant morbidity and mortality worldwide. As the underlying mechanisms of osteoporosis are elucidated, immune dysfunction continues to emerge as a key precipitating factor in multiple bone disease contexts. This review examines recent findings in the osteoimmunology field and their implications for bone disease and for novel future therapeutic approaches to rejuvenate the skeleton. RECENT FINDINGS: T-cells and B-cells have long been recognized to play important roles in the etiology of inflammatory bone disease; however, new findings continue to challenge our understanding of the depth of the immuno-skeletal interface. In this review, we examine recent evidence for new roles of B-cells in oestrogen deficiency bone loss; central actions of interleukin-7 in the cause of T-cell mediated tissue destruction in rheumatoid arthritis; novel RANKL-independent alveolar bone loss in periodontal infection; and a putative role for γδ T-cells in bisphosphonate-associated osteonecrosis of the jaw. Finally, evidence for novel bone anabolic activities mediated through T-cells by the CD28 antagonist CTLA-4Ig and by intermittently administered parathyroid hormone are examined. SUMMARY: As the field of osteoimmunology continues to mature, new interrelationships between immune cells and bone turnover continue to emerge.
Subject(s)
Alveolar Bone Loss/immunology , Arthritis, Rheumatoid/immunology , B-Lymphocytes/metabolism , Bone and Bones/immunology , Mandibular Diseases/immunology , Osteoporosis/immunology , T-Lymphocytes/metabolism , Arthritis, Rheumatoid/metabolism , Arthritis, Rheumatoid/physiopathology , Bone Density Conservation Agents/therapeutic use , Bone and Bones/metabolism , Bone and Bones/physiopathology , Humans , Mandibular Diseases/metabolism , Mandibular Diseases/physiopathology , Osteoclasts/metabolism , Osteoporosis/metabolism , Osteoporosis/physiopathologyABSTRACT
BACKGROUND: Osteocytes are increasingly recognized as significant sources of osteoclast differentiation factor, receptor activator of nuclear factor-κB ligand (RANKL), and osteoblast differentiation inhibitory factor, sclerostin. In this study, RANKL and sclerostin expression of osteocytes is investigated in rats with ligature-induced periodontitis. METHODS: Rats were divided into control and periodontitis groups, and periodontitis was induced by ligature on the mandibular first molars. At 1, 3, 10, and 20 days after ligature, histologic analyses of alveolar bone (AB) and osteoid areas in the molar furcation were performed. The numbers of osteoclasts and RANKL- and sclerostin-positive osteocytes were estimated by tartrate-resistant acid phosphatase staining and immunohistochemistry, respectively. RESULTS: The AB area gradually decreased at day 10 after ligature and increased at day 20. The number of osteoclasts markedly increased at day 3 and then decreased. Conversely, osteoid formation was suppressed up to day 3 and then showed a remarkable increase above control level at day 20. The number of RANKL-positive osteocytes increased at days 1 and 3 and then decreased. Sclerostin-positive osteocytes markedly increased at days 3 and 10 but decreased below control level at day 20. CONCLUSIONS: These results show that AB loss is accompanied by enhanced osteoclast formation and suppressed osteoid formation. Osteocytes express RANKL when osteoclast formation increases, and they express sclerostin when osteoid formation is suppressed. Conversely, osteocytic sclerostin expression decreases when osteoid formation increases. These findings suggest that osteocytes may be important in AB loss via RANKL and sclerostin expression in periodontitis.
Subject(s)
Alveolar Process/chemistry , Bone Morphogenetic Proteins/analysis , Osteocytes/chemistry , Periodontitis/metabolism , RANK Ligand/analysis , Acid Phosphatase/analysis , Alveolar Bone Loss/metabolism , Alveolar Bone Loss/pathology , Alveolar Process/pathology , Animals , Apoptosis/physiology , Bone Matrix/chemistry , Bone Matrix/pathology , Genetic Markers , Isoenzymes/analysis , Leukocytes, Mononuclear/pathology , Male , Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Neutrophils/pathology , Osteoclasts/pathology , Periodontitis/pathology , Random Allocation , Rats , Rats, Inbred F344 , Tartrate-Resistant Acid Phosphatase , Time FactorsABSTRACT
Clinical, histological and immunohystochemical studies of keratocystic tumors were performed showing differences in proliferative activity and MMP-9 expression scores in orthokeratinized and parakeratinised cysts. These histological features are associated with clinical course and may be used as markers for recurrence probability.
Subject(s)
Mandibular Diseases/metabolism , Mandibular Diseases/pathology , Odontogenic Cysts/metabolism , Odontogenic Cysts/pathology , Adult , Biomarkers/metabolism , Humans , Ki-67 Antigen/metabolism , Male , Mandibular Diseases/surgery , Matrix Metalloproteinase 9/metabolism , Odontogenic Cysts/surgery , RecurrenceABSTRACT
The term inflammatory myofibroblastic tumor (IMT) encompasses a diverse group of spindle cell entities that traverses a clinical and histologic spectrum, extending from reactive to benign neoplastic to highly aggressive with malignant inclinations. Head and neck IMTs are rarely seen and comprise less than 5 % of tumors. Here we report a case of a 30 year old male who presented with a rapidly enlarging and extremely painful growth in the right posterior mandible, post extraction. Histopathological examination revealed a highly cellular connective tissue stroma comprised of spindle shaped cells arranged in fascicles, admixed with inflammatory cells, predominantly plasma cells. Apart from routine hematological investigations, serum protein electrophoresis was also performed. The final diagnosis was confirmed by a panel of immunomarkers consisting of MPO, CD34, CD20, CD3, CD23, CD138, SMA and ALK. To the best of our knowledge, this is the third case of oral IMT arising from an extraction socket.
Subject(s)
Granuloma, Plasma Cell/pathology , Mandibular Diseases/pathology , Adult , Biomarkers, Tumor/analysis , Disease Progression , Granuloma, Plasma Cell/metabolism , Humans , Immunohistochemistry , Inflammation/pathology , Male , Mandibular Diseases/metabolismABSTRACT
The purpose of this study was to examine histological alterations on osteoblasts from the alveolar bone of transgenic mice with targeted ablation of osteoctyes. Eighteen weeks-old transgenic mice based on the diphtheria toxin (DT) receptor-mediated cell knockout (TRECK) system were used in these experiments. Mice were injected intraperitoneally with 50 µg/kg of DT in PBS, or only PBS as control. Two weeks after injections, mice were subjected to transcardiac perfusion with 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), and the available alveolar bone was removed for histochemical analyses. Approximately 75% of osteocytes from alveolar bones became apoptotic after DT administration, and most osteocytic lacunae became empty. Osteoblastic numbers and alkaline phosphatase (ALP) activity were markedly reduced at the endosteum of alveolar bone after DT administration compared with the control. Osteoblastic ALP activity in the periodontal ligament region, on the other hand, hardly showed any differences between the two groups even though numbers were reduced in the experiment group. Silver impregnation showed a difference in the distribution of bone canaliculi between the portions near the endosteum and the periodontal ligament: the former appeared regularly arranged in contrast to the latter's irregular distribution. Under transmission electron microscopy (TEM), the osteoblasts in the periodontal ligament showed direct contact with the Sharpey's fibers. Thus, osteoblastic activity was affected by osteocyte ablation in general, but osteoblasts in contact with the periodontal ligament were less affected than endosteal osteoblasts.