Signalling molecules in jaw bones and gingival tissues of patients with Class II and Class III dentofacial deformities

OBJECTIVES: To detect signalling molecule specificities in jaw bone growth zones in skeletal class II and class III patients and compare them to those of a control group. MATERIAL AND METHODS: Twenty skeletal Class II and 20 skeletal Class III patients who underwent orthognathic surgery treatment were in the study group and five skeletal Class I patients who had impacted third molars extracted were in the control group. During the orthognathic surgery, tissue samples were taken from the tuber maxillae, ramus mandibulae anterior and posterior part together with mucosa from the gingival transitory fold in the second molar region of the lower jaw. The samples were stained to detect TGF-β, BMP2/4, FGFR1, VEGF, OC, OP and MMP2 expression. The distributions of these factors were assessed semiquantitatively. RESULTS: We observed significant expression of TGF-β, BMP2/4, OC and OP in the bone tissue of the study group. FGFR1 expression was more pronounced only in mucosa. VEGF and MMP2 were found only in some tissue samples. More apoptotic cells were observed in the bone tissue and soft tissue of the control patients than in those of the skeletal Class II and Class III patients, in which apoptotic cell frequencies were relatively equal. CONCLUSION: From bone tissue in tuber maxillae region the greater TGF-β and BMP2/4 expression is seen in Class III and control groups, comparing to Class II. In ramus mandibulae anterior part the expression of significant factors in bone tissue growth (TGF-β un BMP2/4) is higher in the control group and Class II patients, while in ramus mandibulae posterior part higher expression of TGF-β and BMP2/4 is in Class III patients, comparing to Class II, which indicates to a preserved growth potential in these jaw bone regions. More active bone extracellular matrix protein (osteocalcin and osteopontin) expression in tuber maxillae region both in class II and class III patient groups and different expression in ramus mandibulae anterior part, prove to the bone mineralization and metabolism activity changes, which, perhaps, characterize just these dentofacial deformations.publishersversionPeer reviewe

Expression of gene proteins, interleukins and β-defensin in cleft-affected tissue

OBJECTIVES: The aim of the study was to investigate cleft - affected tissues in children with cleft lip and palate in order to detect appearance of β-defensin, interleukins (IL) and gene proteins. MATERIAL AND METHODS: The study group included 10 patients with cleft lip and palate. During nose surgery tissue samples from bone and cartilage have been taken, and stained with immunohistochemistry for β-defensin 2, IL4, IL6, IL7, IL8, IL10 and MSX1, RYK, PAX9, IRF6 gene proteins. RESULTS: Results showed prominent expression of IL10 (mean value 47.28±4.26 in visual field) followed by IL7 (35.62±11.18) in cartilage of patients, but slightly less pronounced expression of IL8 (30.14±8.74), IL6 (22.52±10.88) and IL4 (14.81±6.94). The expression of β-defensin 2 was prominent (34.52±11.79) and similar to expression of IL7 in tissue samples of cartilage. MSX1, PAX9, RYK and IRF6 (17.67±5.94; 16.14±5.52; 16.57±5.22 and 11.86±4.21) in cartilage was less pronounced than interleukins and β-defensin 2. MSX1 (12.44±3.34), PAX9 (6.89±2.14), RYK (11.0±5.92) and IRF6 (9.1±4.76) gene proteins expression in bone showed mostly rare occurrence of positive structures. CONCLUSIONS: Significant expression of IL7 and β-defensin 2 and IL10 in cartilage proves the prominent immune response in cleft affected hard tissue. Mostly indistinct MSX1, PAX9, RYK and IRF6 gene proteins expression in bone might be an indicator of not complete cellular differentiation, proliferation and migration events in cleft disordered hard tissue.publishersversionPeer reviewe

Osteopontin and osteocalcin in maxilla tissue of skeletal Class III patients

Bone extracellular matrix proteins osteopontin and osteonectin have important function during embryonic osteogenesis and are active in bone remodeling process. Knowledge about bone extracellular matrix proteins explores the mechanisms of craniofacial development and helps to understand better morphopathogenesis of severe dentofacial anomalies. Aim of the current study was to investigate an expression of osteopontin and osteocalcin in maxilla tissue in skeletal Class III patients. The study group included 10 skeletal Class III patients to whom combined orthodontic and surgical treatment was recommended. Samples of tuber maxillae tissue from ten skeletal Class III malocclusion patients were collected due the orthognatic surgery procedure. The routine histological method was followed and samples were stained by hematoxyline-eosine. Tissue samples were stained according the standard immunohistochemical methods and expression of osteocalcin and osteopontin in maxilla tissue was examined. Distribution of these factors was evaluated semiquantitetively by counting the positive structures in visual field. The trabecules of bone contained parallel located collagen fibers and irregular bone mineralization. Connective tissue proliferation in osteon channels was observed. Quantity of cells positive to osteopontin was higher than quantity of cells containing osteocalcin in all patterns extracted of maxilla tissue in patients with skeletal Class III. In few cases osteocalcin positive cells were located near the border with soft tissue - close to osteogenic cells zone, but no osteocalcin was observed deeper in bone. The intensity of osteocalcin expression was variable, and differs individually despite the relatively regular osteopontin distribution in all patterns. There were pronounced osteopontin, but not osteocalcin-containing cells with approximately similar intensity of protein expression in tuber maxillae in skeletal Class III patients.publishersversionPeer reviewe

Specific signaling molecule expressions in the interradicular septum in different age groups

Orthodontic teeth movement is accompanied by the remodeling of alveolar bone, including the interradicular septum. Bone contains three cell types, osteoblasts, osteocytes, and osteoclasts that are in direct contact with all of the cellular elements in the bone marrow. Marrow is the source of both bone-building osteoblasts and bone destroying osteoclasts, and the turnover of bone occurs throughout life. Bone signalling molecules have important functions during osteogenesis, and they are active in the bone remodelling process. Patients involved in orthodontic treatment belong to different age groups: therefore age must be considered as a contributing factor compromising the osteogenetic potential of bone. The aim of the current study was to investigate the specific expression of signalling molecules in the interradicular septum in different age groups. The study group included 17 patients to whom the extraction of teeth was recommended as part of further orthodontic treatment. Patients (9 males and 8 females) - were divided into 3 groups 1st group - 12-14 years old); 2nd group - 15-22 years old; 3rd group - 23 years old or older. Expression of BMP 2/4, TGF-α, IL-1, IL-8, OPG, MMP-1, MMP-2, MMP-8, MMP-9, MMP-13, NGFR, NKpB 105, osteocalcin, and osteopontin in interradicular septum tissues was examined. TUNEL staining was also completed. The distribution of these factors was evaluated semi quantitalively. In the interradicular septum bone structure, the expression levels of osteocalcin, osteoprotegerin, matrix metalloproteinases 8 and 9, and nuclear factor kappa B were determined in all samples. TUNEL staining was also done. Age related decreases in the mean values of signalling factors and the number of apoptotic cells were statistically significant. Specific to interradicular septum osteoblasts and osteoclasts factors include osteoprotegerin, osteocalcin, matrix metalloproteinase 8, matrix metallproteniase 9, and nuclear factors kappa B. The mean expression levels of these proteins and the mean TUNEL staining statistically significantly decreased with age. This is preliminary study and more patients are necessary for more precise statistical analysis in the future.publishersversionPeer reviewe

Correlation of temporomandibular joint clinical signs with cone beam computed tomography radiologic features in juvenile idiopathic arthritis patients

OBJECTIVE: Patients with juvenile idiopathic arthritis (JIA) have a high risk of temporomandibular joint (TMJ) involvement. Early detection of osseous destruction of the TMJ that can be seen radiographically is vital to provide appropriate treatment before significant craniofacial deformities and problems with occlusion arise. The aim of study was to evaluate whether there is a correlation between the clinical signs and cone beam computed tomography (CBCT) radiologic features of TMJ in patients with JIA. MATERIAL AND METHODS: Study group consisted of 65 patients (46 females and 19 males) aged 10 to 17 years with a confirmed JIA diagnosis and mean disease duration 2 years 9 month, all patients underwent a clinical examination of the TMJ and masticatory muscles as well as a radiological assessment of the TMJ osseous structures by CBCT. RESULTS: Majority of the patients' study population experienced 2 or 3 clinical signs with mean number 2.1 (standard deviation (SD) =1.00) and 3-5 radiological features related to the TMJ destruction with mean number 4.9 (SD=1.96). Statistically significant weak correlation only between pain and condyle surface flattening (Spearman`s Rho test (rho) =0.396; p value (p) =0.001) was found. No correlation between number of clinical signs and radiological features was found. CONCLUSIONS: There was no conclusive evidence found regarding correlation between TMJ clinical signs and radiological features of osseous destruction in patients with JIA - only weak correlation between pain and condyle surface flattening was observed. The number of clinical TMJ signs does not correlate with number of radiological features. For the clinical decision both clinical examination and CBCT would be useful in the early detection of osseous destruction of the TMJ in JIA patients.publishersversionPeer reviewe

The relationship between mandibular rotation and osseous structure of the TMJ in pre-surgery orthognathic patients : A cone beam CT study

The aim of the study was to assess the relationship between vertical skeletal pattern in terms of mandibular rotation and osseous structural changes of the TMJ in pre- surgical orthognatic patients. TMJ skeletal morphology was evaluated in cone beam computer tomography images of 117 consecutive patients with Class II and Class III dentofacial deformities according to the research diagnostic criteria of the osseous components of the TMJ related to the maxillary-mandibular plane (MM) angle. The distribution of the number and percentage of joints with structural changes in Class II was markedly different in groups divided according to the MM angle. Statistically significant increase was found in the percentage of TMJ's with osseous changes separately for each side, i.e., right (p=0.001), left (p=0.04) and both together (p=0.0001), in the Class II patient group, an increased MM angle indicated backward rotation of the mandible. In Class III patients, there were no statistically significant differences in the number of joints with TMJ structural changes. The presence of mentioned changes was asymmetrical between the left and right joints in both the Class II and Class III patient groups. In conclusion, structural changes in the osseous parts of the TMJ are more common in patients with Class II skeletal dentofacial deformities with backward rotation of the mandible than in Class III pre-surgery orthognathic patients.publishersversionPeer reviewe

Growth factors, apoptotic cells and barx1 gene in bone and soft tissue of skeletal class III patients

Growth factors and growth stimulating genes are main signaling molecules for growth and development in ante- and postnatal period involved in cellular proliferation, differentiation and morphogenesis of tissues and organs during embryogenesis, postnatal growth and adulthood. The aim of this study was to evaluate TGF-beta (transforming growth factor-beta), BMP2/4 (bone morphogenetic protein 2/4), FGFR1 (fibroblast growth factor receptor one), barx1 gene and apoptosis from tissue samples of oro-maxillo-facial region in skeletal class III patients to reveal possible morphopathogenesis of severe skeletal anomalies. The study group included 9 patients with skeletal class III malocclusion. During orthognatic surgery tissue samples from tuber maxillae, ramus mandibulae anterior and posterior part, as well as gingiva from the lower jaw in region of second molar have been taken. Samples were stained with immunohistochemistry for TGF-beta, BMP2/4, FGFR1, apoptosis and barx1 gene. We used also the routine histological staining with haematoxyline and eosine. In tuber maxillae, ramus mandibulae anterior and posterior part staining for TGF-beta was the most relevant. Also BMP2/4, FGFR1 and barx1 showed the highest mean number of positive cells in tuber maxillae. Barx1 was equally expressed in ramus mandibulae, but BMP2/4 and FGFR1 mainly stained its posterior part cells. Apoptosis mostly affected ramus mandibulae anterior part. CONCLUSIONS: We suggest about more active stimulation of bone growth in tuber maxilla whereas ramus mandibulae. Apoptosis mainly affects ramus mandibulae anterior part that possibly connects to the lower expression of growth stimulating factors and may indicate lower bone remodelation ability.publishersversionPeer reviewe

Radiologic features of temporomandibular joint osseous structures in children with juvenile idiopathic arthritis. Cone beam computed tomography study

INTRODUCTION: Patients with juvenile idiopathic arthritis (JIA) have a high risk of temporomandibular joint (TMJ) involvement. Lesions in the TMJ appear early in the course of this disease. Evaluating the structure of the TMJ in JIA patients using cone beam computed tomography (CBCT) provides an understanding of the typical radiologic features of morphological change in TMJs of JIA patients. This study aims to report these features as seen in CBCT and thus comparing them with the features observed in a control group within the same age group and in females and males. MATERIALS AND METHODS: Cross-sectional observational study whereby CBCTs of 65 (130 joints) patients with a confirmed JIA diagnosis and 30 (60 joints) control group - patients without JIA upto the age of 17. Structural radiologic features of the joint's hard tissues were assessed according to the research diagnostic criteria for temporomandibular disorders as developed by Ahmad et al. RESULTS: The radiologic features of the osseous structures of the TMJ occurred asymmetrically between the right and left sides when compared in the JIA and control groups. The most prevalent feature in the JIA group is condyle surface flattening for both sides. Condyle surface erosion and osteophyte were also frequent and occurred with high statistical significance in both males and females. CONCLUSIONS: TMJ destruction features observed in CBCT images were prevalent in the JIA group and occurred infrequently in the control group.publishersversionPeer reviewe

Standardizing Terminology and Assessment for Orofacial Conditions in Juvenile Idiopathic Arthritis : International, Multidisciplinary Consensus-based Recommendations

Objective. To propose multidisciplinary, consensus-based, standardization of operational terminology and method of assessment for temporomandibular joint (TMJ) involvement in juvenile idiopathic arthritis (JIA). Methods. Using a sequential expert group–defined terminology and methods-of-assessment approach by (1) establishment of task force, (2) item generation, (3) working group consensus, (4) external expert content validity testing, and (5) multidisciplinary group of experts final Delphi survey consensus. Results. Seven standardized operational terms were defined: TMJ arthritis, TMJ involvement, TMJ arthritis management, dentofacial deformity, TMJ deformity, TMJ symptoms, and TMJ dysfunction. Conclusion. Definition of 7 operational standardized terms provides an optimal platform for communication across healthcare providers involved in JIA-TMJ arthritis management.publishersversionPeer reviewe