Cementocyte cell death occurs in rat cellular cementum during orthodontic tooth movement

Objective:To clarify the mechanism of root resorption during orthodontic treatment, we examined cementocyte cell death and root resorption in the cellular cementum on the pressure side during experimental tooth movement.Materials and Methods:Using 8-week-old male Wistar rats, the right first molar was pushed mesiobuccally with a force of 40 g by a Ni-Ti alloy wire while the contralateral first molar was used as a control. Localization and number of cleaved caspase-3-positive and single-stranded DNA (ssDNA) - positive cells were evaluated using dual-label immunohistochemistry with anticleaved caspase-3 and anti-ssDNA antibodies. In addition, tartrate-resistant acid phosphatase (TRAP)-positive cells in the cellular cementum were evaluated using TRAP histochemical staining.Results:Caspase-3- and ssDNA-positive cells appeared at 12 hours, but were restricted to the compressed periodontal ligament (PDL) and not the cellular cementum. Cleaved caspase-3-positive cementocytes were observed in the cellular cementum adjacent to the compressed PDL on day 1. From days 2 to 4, the number of caspase-3- and ssDNA-positive cementocytes increased. TRAP-positive cells appeared on the cellular cementum at the periphery of the hyalinized tissue on day 7, and resorption progressed into the broad surface of the cementum by day 14.Conclusion:Cementocytes adjacent to the hyalinized tissue underwent apoptotic cell death during orthodontic tooth movement, which might have been associated with subsequent root resorption.     

VDR deficiency affects alveolar bone and cementum apposition in mice

Objective

To compare the mineralisation density (MD), morphology and histology of alveolar bone and cementum amongst VDR +/+, VDR −/−, and VDR −/− groups supplemented with a diet TD 96348, containing 20% lactose, 2.0% calcium and 1.25% phosphorous.

Methods

Four groups of mice (6 mice/group) were identified by genotyping: VDR +/+ mice (VDR wild type), VDR −/− mice (VDR deficient), VDR −/− offsprings derived from VDR −/− parents receiving a supplemental diet (early rescued), and VDR −/− mice fed with a supplemental diet beginning at age one month (late rescued). All mice were sacrificed at age 70.5 days. Micro-CT was used to compare MD and morphology of alveolar bone and cementum. H–E and Toluidine blue staining was used to examine the ultrastructure of the alveolar bone and cementum at matched locations.

Results

In VDR −/− group, alveolar bone and cementum failed to mineralise normally. Early rescue increased MD of alveolar bone in VDR −/− mice with excessive alveolar bone formation, but which not observed in late rescue group. MD and morphology of cementum–dentine complex in both early and late rescue groups were comparable with VDR +/+ group when feeding with high-calcium rescue diet.

Conclusions

VDR affects alveolar bone mineralisation and formation systemically and locally. However, cementum apposition and mineralisation is mainly regulated by calcium concentrations in serum.     

Expression and distribution of SIBLING proteins in the predentin/dentin and mandible of hyp mice

Oral Diseases (2010) 16 , 453–464 Objectives: Human X‐linked hypophosphatemia (XLH) and its murine homologue, Hyp are caused by inactivating mutations in PHEX gene. The protein encoded by PHEX gene is an endopeptidase whose physiological substrate(s) has not been identified. Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP), two members of the Small Integrin‐Binding LIgand, N‐linked Glycoprotein (SIBLING) family are proteolytically processed. It has been speculated that PHEX endopeptidase may be responsible for the proteolytic cleavage of DMP1 and DSPP. To test this hypothesis and to analyse the distribution of SIBLING proteins in the predentin/dentin complex and mandible of Hyp mice, we compared the expression of four SIBLING proteins, DMP1, DSPP, bone sialoprotein (BSP) and osteopontin (OPN) between Hyp and wild‐type mice. Methods: These SIBLING proteins were analysed by protein chemistry and immunohistochemistry. Results: (1) Dentin matrix protein 1 and DSPP fragments are present in the extracts of Hyp predentin/dentin and bone; (2) the level of DMP1 proteoglycan form, BSP and OPN is elevated in the Hyp bone. Conclusions: The PHEX protein is not the enzyme responsible for the proteolytic processing of DMP1 and DSPP. The altered distribution of SIBLING proteins may be involved in the pathogenesis of bone and dentin defects in Hyp and XLH.     

Disturbed tooth eruption in osteopetrotic (op/op) mice: histopathogenesis of tooth malformation and odontomas

BACKGROUND: Odontoma-like structures are formed in the jaw bone of osteopetrotic (op/op) mice, which have a congenital deficiency in osteoclastic differentiation due to the absence of functional macrophage colony-stimulating factor (M-CSF). METHODS: To clarify the histopathogenesis of tooth malformation and odontoma-like structures, a 2-year postnatal process of development of the op/op mandibular incisor was examined radiologically and histologically. At the same time, extracellular matrix (ECM) remodeling around tooth germs was analyzed immunohistochemically. RESULTS: Abnormal forms of op/op tooth germ were noticeable even at 3 days after birth on a radiogram. Histologically, op/op mice were clearly distinguished by the disappearance of dental follicular space at 3 days. With aging, bone trabeculae, which were not remodeled, penetrated into op/op tooth germs and divided them into several daughter germs, which were recognized as odontomas. In mandibular incisor bodies, the immature ECM components, such as heparan sulfate proteoglycan and tenascin, were preserved diffusely in the dental papilla/pulp, which indicates that maturation of the stroma does not take place in op/op mandibular incisors. CONCLUSION: The observation suggests that the disturbed morphogenesis of op/op tooth germs is functionally explained by the disordered immunolocalization of ECM molecules, and that the dental follicular space is essential for normal tooth development because it prevents bone penetration into the tooth germs.     

Comparative study of gene expression during tooth eruption and orthodontic tooth movement in mice

Objective:  The aim of this study was to understand tooth eruption by comparing the gene expression during tooth eruption and orthodontic tooth movement (OTM).
Materials and methods:  Orthodontic force was applied on maxillary molars for 2, 4, 7 and 14 days to study tooth movement. Mice at PN 0, 7, 10, 15 and 21 were fixed to observe tooth eruption. Comparative study of two procedures was assessed by haematoxylin and eosin, tartrate-resistant acid phosphatase staining and in situ hybridization for matrix metalloproteinase ( Mmp ) 2 , 13 , bone sialoprotein ( Bsp ) and osteocalcin ( Ocn ).
Results:  Tartrate-resistant acid phosphatase activity and expression of Mmp2 , 13 were obviously detectable in the compression region during OTM. They were also identified in the occlusal and apical region of alveolar bone during tooth eruption. Strong expression of Bsp and Ocn was detectable at the tension side during OTM. These genes were also expressed in the inner lateral region of alveolar bone adjacent to the tooth, but absent in the inner surface of the occlusal and root apical regions during tooth eruption.
Conclusion:  The process of alveolar bone metabolism during developmental eruption and OTM shares the same mechanism. Internal force, as the orthodontic force for OTM, may be initiating factor for tooth eruption.     

Osteonectin RNA and collagen α1(I) RNA in the developing rat maxilla

The sequence of rat osteonectin mRNA was determined. Comparison with osteonectin (ON) mRNA sequences of other vertebrates showed a great similarity, but a stretch of codons deviated with respect to another rat strain, suggesting the possibility of two ON variants in rats. Northern blots exhibited one band of ON mRNA only. The expression of ON and collagen α1(I) RNA in tissues of the developing rat maxilla was studied by in situ hybridization. Both ON and collagen α1(I) RNA were observed concomitantly in osteoblasts, starting from the onset of bone formation at day 17 post coitum through the oldest age examined, day 20 after birth. A strict co-expression of the two sequences was also observed in odontoblasts as well as in fibroblasts of the periodontal ligament. In ameloblasts, neither ON nor collagen α1(I) RNA was detected under stringent hybridization conditions, but lower stringency led to an ON signal. Considering that ON is a secretory protein and the high stability of the ON mRNA, the co-expression of collagen α1(I) and ON RNA sequences in matrix-forming cells provided evidence that ON is a substantial component of collagen matrices.     

颞下颌关节盘摘除术关节区组织病理学变化的实验研究

目的 建立兔颞下颌关节盘摘除术实验动物模型,研究关节盘摘除术早期,关节区组织形态学变化.方法 用10只新西兰大白兔,实验组8只行双侧关节盘摘除术;2只为正常对照组.术后1周、2周、4周、10周各处死2只,切取关节组织,进行组织病理学观察.结果 髁突及关节结节关节软骨连续性破坏,功能区关节软骨下骨组织直接暴露于关节腔内,非功能区则软骨细胞各层增生明显,表面纤维层增生变厚,呈现出纤维性粘连样改变.暴露在关节腔部分的骨组织表面致密,髓腔内的骨小梁吸收,伴微小囊肿形成.髁突关节软骨、骨组织及滑膜出现早期骨关节炎样改变.结论 兔关节盘摘除术后早期,关节区组织表现为骨关节炎样改变,不是适应性改变.     

Hypertension favors the inflammatory process in rats with experimentally induced periodontitis

Bonato CF, do‐Amaral CCF, Belini L, Salzedas LMP, Oliveira SHP. Hypertension favors the inflammatory process in rats with experimentally induced periodontitis. J Periodont Res 2012; 47: 783–792. © 2012 John Wiley & Sons A/S Background and Objective: Cardiovascular diseases are significantly correlated with chronic periodontitis. The aim of this study was to evaluate bone‐loss level, neutrophil migration, CXCL2/CINC‐2α, CXCL5/LIX, CCL20/MIP‐3α and tumor necrosis factor‐α (TNF‐α) production, inducible nitric oxide synthase (iNOS) expression and C‐reactive protein (CRP) release in spontaneously hypertensive rats (SHRs) and normotensive (WTK) rats after experimental induction of periodontal disease. Material and Methods: Periodontitis was induced by placement of silk yarn ligatures around the first molar counterparts. The levels of CRP, CCL20/MIP‐3α and CXCL5/LIX were evaluated in the peripheral blood, and bone‐loss level, neutrophil recruitment, the production of myeloperoxidase, CXCL2, CXCL5, CCL20 and TNF‐α, and the expression of iNOS were evaluated in the gingival tissue. Histological sections were taken to evaluate and measure bone resorption and neutrophil recruitment in the furcation region. Results: Rats with periodontitis had alveolar bone resorption. SHRs with periodontitis showed marked bone loss and increased neutrophil infiltration in comparison with WTK rats. SHRs with periodontitis showed increased levels of TNF‐α and CXCL2, and a slight tendency for increased levels of CXCL5, in the gingival tissue but no increase in the level of CCL20. In SHRs, even without periodontitis, the levels of TNF‐α, CXCL2, CXCL5 and CCL20 showed a slight tendency to increase. In the WTK rats, TNF‐α, CXCL2 and CXCL5 levels were increased with periodontitis, but the level of CCL20 was not. iNOS was expressed in the gingival tissue of WTK rats and SHRs with periodontitis; however, SHRs appeared to express a higher level of iNOS than did WKT rats. The CRP level was elevated in both types of rats with periodontitis; however, the CRP level was higher in SHRs with periodontitis than in WTK rats with periodontitis. Conclusion: In SHRs, the hypertensive condition per se seems to favor the inflammatory processes that become potentiated with periodontitis, when compared with WKT rats.     

Cloning of hamster osteopontin and expression distribution in normal tissues and experimentally induced oral squamous-cell carcinoma

Osteopontin (OPN) is a non-collagenous extracellular matrix (ECM) protein expressed and secreted by several human cancers. This study investigated the expression pattern of OPN during development of oral squamous-cell carcinoma by using 7,12-dimethylbenz[a]anthracene (DMBA)-induced squamous-cell carcinomas in buccal pouch of syrian golden hamsters. We first identified the hamster OPN cDNA sequence by screening of a hamster calvariae cDNA library with a rat OPN cDNA probe. The resulting 1,449 bp of hamster OPN cDNA led to a deduced protein sequence of 305 amino acids containing several putative binding sites to integrins, CD44 receptors, calcium ions and hydroxyapatite, as well as multiple sites for phosphorylation, glycosylation and sulphation. Hamster OPN cDNA was then used as a probe to analyze the expression of OPN mRNA by Northern blot and in situ hybridization analyses of normal and malignant tissues. OPN mRNA was detected in several non-mineralized tissues as well as in mineralized tissues, but was not present in normal hamster buccal epithelium. DMBA-treated hamster buccal pouches expressed OPN mRNA as early as 4 weeks and displayed the highest level of expression at 15 weeks. The specimens treated with DMBA for 15 weeks exhibited histological features of squamous-cell carcinoma, presented microcrystalline deposits and showed OPN expression associated with malignant epithelium and tumor-associated macrophages. To summarize, our results suggest that buccal-pouch carcinogenesis of Syrian golden hamster may constitute an excellent experimental model to study the mechanisms by which OPN is associated with oral cancer pathogenesis, and to validate OPN-based therapeutic approaches to ameliorate oral cancer progression and metastasis.     

颞下颌关节盘前移位关节区组织病理变化的实验研究

目的通过建立关节盘前移位实验动物模型的方法,来研究人类关节盘前移位病变早期阶段的发展过程和病理学改变。方法１２只新西兰大白兔左颞下颌关节经实验诱导为关节盘前移位模型,右侧为手术对照组。于术后２４ｈ、１周、２周、３周、４周、１０周各处死２只,切取关节组织,ＨＥ染色,光镜观察。结果早期：关节结节、髁突关节软骨增生明显,髁突软骨下出血,骨组织内血管网消失;后期：髁突关节软骨、骨组织及滑膜出现骨关节炎（ＯＡ）样改变。结论实验诱导关节盘前移后关节区的病理学变化与人类相似;关节盘前移位关节区的创伤可引起关节骨关节炎样改变。     

Functional changes in the temporomandibular joint mechanoreceptors associated with experimentally induced condylar resorption in rats

ObjectivesTo evaluate the influence of experimentally induced progressive condylar resorption (PCR) on temporomandibular joint (TMJ) mechanoreception.Materials and MethodsTwenty 13-week-old male albino Wistar rats were divided equally into control and PCR groups. A compressive force was loaded on the left TMJ of PCR group rats to induce condylar resorption. Single-unit activities of TMJ mechanoreceptors were also induced through passive jaw movement. Recording was performed for the left Gasserian ganglion at 3 days and 1 week after the establishment of PCR group. The effects of PCR on TMJ units were assessed by measuring the firing threshold, maximum instantaneous firing frequency, and average firing frequency.ResultsCompared with the control group, there were no significant differences in the firing threshold of the PCR group after 3 days. The thresholds were significantly higher 1 week after compressive force loading on the condyle. The maximum instantaneous firing frequencies and the average firing frequencies showed no significant differences after 3 days. However, these were significantly lower 1 week after compressive force loading.ConclusionsThe findings suggest that compressive force loading on the condyle may influence the function of TMJ mechanoreceptors.