Characterization of Mandibular Bones in Senile Osteoporotic Mice

At present, little is known about the age-related changes in jaw bones. The aim of this study was to characterize the mandibles of 6 month-old senile osteoporotic mice, SAMP6, and compare with those of age-matched controls, SAMR1. In comparison to SAMR1, SAMP6 showed thinner cortical bone, lower bone volume, and poorly organized collagen matrix. The collagen fibril diameter in SAMP6 was significantly smaller than that of SAMR1. In SAMP6 both collagen content and cross-links were lower than those of SAMR1, but the ratio of the major mature cross-link (pyridinoline) to its precursor reducible cross-link (dehydrodihydroxylysinonorleucine/its ketoamine) was higher in comparison to SAMR1. In addition, the extent of lysine hydroxylation of collagen was higher in SAMP6 than that of SAMR1. These results indicate that not only the quantity of collagen but also its quality are altered in SAMP6 and may result in the age-associated osteoporotic defects of mandibles.     

Long-term effects of various cleaning methods on polypropylene/ethylene copolymer retainer material

Objectives:To evaluate long-term light transmittance, surface roughness, and flexural modulus of polypropylene/ethylene copolymer retainer material after exposure to different cleaning methods.Materials and Methods:Standardized polypropylene/ethylene copolymer retainer specimens (n = 70, 50.8 mm × 12.7 mm × 1.0 mm) were subjected to seven chemical cleaning solutions: Invisalign cleaning crystals, Retainer Brite, Polident, Listerine mouthwash, 2.5% acetic acid, 0.6% NaClO, and 3% H₂O₂ for 6 months. The specimens were exposed to the different solutions twice a week for 15 minutes or according to manufacturer''s instructions, then stored in artificial saliva at 37°C. Another group of specimens (n = 10) were brushed with a standardized toothbrushing machine for 2 minutes twice a week. At baseline and 6 months, light transmittance, surface roughness, and flexural modulus of the specimens were quantified using spectrophotometry, profilometry and three-point bend testing, respectively. Qualitative analysis was performed using a scanning electron microscope (SEM). Statistical analyses were performed at a significance level of .05.Results:The results showed that light transmittance decreased significantly from baseline for all cleaning methods at 6 months. For an individual method, no significant differences were observed between specimens at baseline and 6 months in surface roughness and flexural modulus. No discernible differences in surface features were observed on SEM images.Conclusions:The results indicate that different cleaning methods affect the long-term light transmittance of the studied polypropylene/ethylene copolymer retainer material. However, for an individual cleaning method, no significant differences were shown for surface roughness or flexural modulus values at 6-months compared to baseline.     

Changes in matrix phosphorylation during bovine dentin development

Phosphorylation of the organic matrix proteins of dentin is important for the initiation of mineralization, but its relevance in later mineralization stages is controversial. The objective of this study was to analyze changes in the total matrix phosphate content during dentin development and to identify their origin. Amino acid and total matrix phosphate analyses of microdissected developing mantle and circumpulpal fetal bovine dentin specimens were performed. The amino acid composition showed few changes during mantle and circumpulpal dentin maturation. However, the total matrix phosphate content showed a significant, positive correlation with tissue maturation in both mantle and circumpulpal dentin, with a two- and a three-fold increase, respectively, being observed. The data indicate that changes occur in the pattern of phosphorylation of matrix proteins during dentin maturation, which we suggest may play a functional role in later stages of tooth mineralization.     

Characterization of mandibular bones in senile osteoporotic mice

At present, little is known about the age-related changes in jaw bones. The aim of this study was to characterize the mandibles of 6 month-old senile osteoporotic mice, SAMP6, and compare with those of age-matched controls, SAMR1. In comparison to SAMR1, SAMP6 showed thinner cortical bone, lower bone volume, and poorly organized collagen matrix. The collagen fibril diameter in SAMP6 was significantly smaller than that of SAMR1. In SAMP6 both collagen content and cross-links were lower than those of SAMR1, but the ratio of the major mature cross-link (pyridinoline) to its precursor reducible cross-link (dehydrodihydroxylysinonorleucine/its ketoamine) was higher in comparison to SAMR1. In addition, the extent of lysine hydroxylation of collagen was higher in SAMP6 than that of SAMR1. These results indicate that not only the quantity of collagen but also its quality are altered in SAMP6 and may result in the age-associated osteoporotic defects of mandibles.     

Decreased expression of lysyl hydroxylase 2 (LH2) in skin fibroblasts from three Ehlers-Danlos patients does not result from mutations in either the coding or proximal promoter region of the LH2 gene

The Ehlers-Danlos syndromes (EDS) are a heterogeneous group of inherited connective tissue disorders characterized by tissue fragility, hyperelasticity of the skin and joint hypermobility. This phenotype, accompanied by kyphoscoliosis and/or ocular fragility, is present in patients with the autosomal recessive type VI form of EDS. These patients have significantly decreased levels of lysyl hydroxylase (LH) activity, due to mutations in the LH1 gene. LH hydroxylates specific lysine residues in the collagen molecule that are precursors for the formation of cross-links which provide collagen with its tensile strength. No disorder has been directly linked to decreased expression of LH2 and LH3, two other isoforms of LH. This study describes 3 patients with mixed phenotypes of EDS, who have significantly decreased mRNAs for LH2, but normal levels of LH1 and LH3 mRNAs, in their skin fibroblasts. In contrast to the effect of LH1 deficiency in EDS VI patients, the decreased expression of LH2 does not affect LH activity, bifunctional collagen cross-links (measured after reduction as dihydroxylysinonorleucine (DHLNL) and hydroxylysinonorleucine (HLNL)), or helical lysine hydroxylation in these cell lines. Sequence analysis of full length LH2 cDNAs and 1kb of the promoter region of LH2 does not show mutations that could explain the decreased expression of LH2. These results suggest that the deficiency of LH2 in these fibroblasts may be caused by changes in other factors required for the expression of LH2.     

Ameloblastin regulates cell attachment and proliferation through RhoA and p27

The matrix adhesion protein ameloblastin (AMBN) is one of the unique components of the mineralizing matrix of bones and teeth. Here we focused on two types of cells expressing AMBN - mouse dental follicle cells (mDF) and mouse periodontal ligament cells (mPDL) - to decipher AMBN function in developing dental, periodontal, and bone tissues. To test AMBN function, cell culture dishes of mDF and mPDL were exposed to either full-length or C-terminal (amino acids 137-407) recombinant Ambn protein. Alternatively, cells were subjected to transient transfection using an Ambn-small hairpin (sh) RNA vector. Our cell culture studies documented that dishes coated with full-length AMBN promoted the attachment of mPDL and mDF cells as early as 1 h after seeding. In order to identify potential intermediaries that might aid the effect of AMBN on adhesion, RhoA expression levels in AMBN-coated and uncoated control dishes were assessed. These studies indicated that AMBN induced RhoA expression 4 h after seeding, especially in mPDL cells. After 4 h of culture, the cell cycle inhibitor p27 was also up-regulated. In addition, exogenous AMBN and its C-terminal fragment reduced the proliferation of mDF and mPDL. Finally, transient transfection of mDF and mPDL cells with the Ambn-shRNA vector resulted in the down-regulation of p27 in mPDL cells. Together, these data indicate that AMBN affects cell adhesion via RhoA and cell cycle progression through p27.     

A novel proteolytic processing of prolysyl oxidase

Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residues Gly(162) and Asp(163) (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity, and mass spectrometry. One form was identified as a well-characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX resulting from the cleavage at the carboxy terminus of Arg(192). The truncated form of LOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX.     

Collagen dysregulation in the dermis of the Sagg/+ mouse: a loose skin model

The Sagg/+ mouse is an ethylnitrosourea-derived mutant with a dermal phenotype similar to some of the subtypes of Ehlers-Danlos syndrome (EDS) and cutis laxa. The dermis of the Sagg/+ mouse has less dense and more disorganized collagen fibers compared to controls. The size of extracted Type I dermal collagen was the same as that observed in normal skin; however, more collagen could be extracted from Sagg/+ skin, which also showed decreased collagen content and decreased steady-state levels of alpha1(I), alpha2(I), alpha1(V), and alpha2(V) procollagen mRNAs. The biomechanical properties of Sagg/+ skin were significantly decreased relative to normal skin. However, there were no significant differences in the quantities of the major collagen cross-links, that is, dehydrohydroxylysinonorleucine and dehydrohistidinohydroxymerodesmosine between Sagg/+ and normal skin. Electron microscopic evaluation of Sagg/+ skin indicated that the mutation interferes with the proper formation of collagen fibrils and the data are consistent with a mutation in Type V collagen leading to haploinsufficiency with the formation of two sub-populations of collagen fibrils, one normal and one with irregular shape and a larger diameter. Further study of this novel mutation will allow the identification of new mechanisms involved in the regulation of normal and pathologic collagen gene expression.     

Cone-beam computed tomography assessment of maxillary anterior alveolar bone remodelling in extraction and non-extraction orthodontic cases using stable extra-alveolar reference

Objective

To explore alveolar cortical positional change in response to tooth movement in extraction and non-extraction orthodontic cases, using cone-beam computed tomography (CBCT) and stable extra-alveolar references.

Materials and Methods

The pre-treatment (T1) and post-treatment (T2) CBCT scans of 25 extraction (EXT) and matched 25 non-extraction (Non-EXT) orthodontic cases were imported into Dolphin Imaging 3D, and oriented uniformly. Sagittal and axial CBCT cross-sections were traced using customized software-generated guides. The displacement of teeth and alveolar bone cortices were automatically measured using the palatal plane (PP) and the line perpendicular to PP and passing Sella as reference. Intra- and inter-group differences between T1 and T2 were analysed. Subjects were also superimposed three-dimensionally using Geomagic Control X for qualitative analysis of cortical remodelling.

Results

The EXT group showed incisor retraction, while the Non-EXT group exhibited statistically significant incisor anterior tipping (P < .05). In EXT, both the labial and palatal cortices are resorbed. Non-EXT showed labial cortex anterior modelling, and statistically significant palatal cortex resorption (P < .05). In both groups, statistically significant decrease in total and palatal alveolar widths, increase in labial widths, and palatal dehiscence were observed. Comparatively, EXT showed significantly more incisal total and palatal width decrease and palatal vertical bone loss.

Conclusion

Labial cortical remodelling was shown to follow anterior tooth movement, but the palatal cortical response to incisor retraction and labial cortical remodelling in general remained inconclusive. Narrowing of the alveolar housing and palatal dehiscence were observed regardless of extraction following orthodontic treatment.