Effects of Lanthanum on Bone Resorbing Activity of Rabbit Mature Osteoclasts Co-Cultured with Osteoblasts

The effects of lanthanum (Ⅲ) on the bone resorbing activity of rabbit mature osteoclasts (OCs) in the presence of osteoblasts (OBs) were studied in vitro by measuring the number and area of absorption pits. La(Ⅲ) at concentrations ranging from 1.00×10-5 to 1.00×10-8 mol·L-1 show no effect on mature OC number (P＞0.05). In the OC-OB co-culture systems without La(Ⅲ), osteoblasts alone did not influence the pit number and area whether the two kinds of cells were in contact or not (P＞0.05). Under the OC-OB not-in-contact condition, the effect of La(Ⅲ) on the bone-resorbing activity of OCs was similar to that of La(Ⅲ) in the absence of OBs (P＞0.05). However, while OCs were in direct contact with OBs, the inhibitory effects of La(Ⅲ) on OCs' bone-resorbing activity decreased at the concentrations of 1.00×10-5, 1.00×10-6 and 1.00×10-7 mol·L-1, and the promotion effects increased at 1.00×10-8 mol·L-1 (P＜0.05). The results suggest that direct cell-cell contact between OC and OB be essential for OBs to play their role in regulating the response of OCs to La(Ⅲ).     

Effects of Lanthanum on Formation and Bone-Resorbing Activity of Osteoclast-Like Cells

The effect of La3 on formation of osteoclast-like cells in rabbit bone marrow cells induced by 1,25-dihydroxyvitamin D3 and their bone-resorbing activity was evaluated by counting the number of tartrate resistant-acid phosphatase-positive ［TRAP( )］ multi-nucleated cells and measuring the number and surface area of bone resorption pits with photomicrography and image analysis. The formation and morphological characteristics of osteoclast-like cells and bone resorption pits were observed under a phase contrast inverted microscope. La3 promotes the formation of osteoclast-like cells at the concentration of 1.00×10-8mol·L-1 compared with the control group(P<0.01), whereas no significant change in cell number is observed at higher concentrations(1.00×10-5, 1.00×10-6 and 1.00×10-7 mol·L-1)(P>0.05). La3 at the concentration of 1.00×10-8mol·L-1 also increases the number and surface area of the resorption pits(P<0.01), but inhibits the bone-resorbing activity dose-dependently(P<0.01)at higher concentrations(1.00×10-5, 1.00×10-6 and 1.00×10-7 mol·L-1). These findings suggest that La3 may promote or inhibit the formation and bone-resorbing activity of osteoclast-like cells depending on its concentration.     

Quantitative and reliable in vitro method combining scanning electron microscopy and image analysis for the screening of osteotropic modulators

The increased generation and up-regulated activity of bone resorbing cells (osteoclasts) play a part in the impairment of bone remodeling in many bone diseases. Numerous drugs (bisphosphonates, calcitonin, selective estrogen receptor modulators) have been proposed to inhibit this increased osteoclastic activity. In this report, we describe a pit resorption assay quantified by scanning electron microscopy coupled with image analysis. Total rabbit bone cells with large numbers of osteoclasts were cultured on dentin slices. The whole surface of the dentin slice was scanned and both the number of resorption pits and the total resorbed surface area were measured. Resorption pits appeared at 48 h and increased gradually up to 96 h. Despite the observation of a strong correlation between the total resorption area and the number of pits, we suggest that area measurement is the most relevant marker for osteoclastic activity. Osteotropic factors stimulating or inhibiting osteoclastic activity were used to test the variations in resorption activity as measured with our method. This reproducible and sensitive quantitative method is a valuable tool for screening for osteoclastic inhibitors and, more generally, for investigating bone modulators.     

Calcitonin gene-related peptide (CGRP)-containing nerve fibers in bone tissue and their involvement in bone remodeling

Bone remodeling is a process of bone renewal accomplished by osteoclastic bone resorption and osteoblastic bone formation. These two activities are regulated by systemic hormones and by local cytokines and growth factors. Moreover, the nervous system and certain neuropeptides seem to be involved in regulation of bone remodeling. In this paper, we focus on the distribution of CGRP-containing nerve fibers and their dynamics, and discuss the role of these fibers as a possible mechanism for nervous system involvement in regulation of bone remodeling. CGRP-immunoreactive nerve fibers are widely distributed in bone tissue, such as periosteum and bone marrow, and show apparent regional distribution with different densities. They are often associated with blood vessels and show a beaded appearance. The wide distribution of CGRP-immunoreactive nerve fibers in bone tissue and the changes in distribution during bone development and regeneration suggest the involvement of these fibers in bone remodeling. The effect of CGRP on bone remodeling could partly be through its action on blood vessels, thereby regulating local blood flow. Moreover, in vitro biochemical data and the localization of CGRP-immunoreactive nerve fibers in the vicinity of bone cells suggest that they are directly involved in local regulation of bone remodeling by elevating the concentration of CGRP in the microenvironment around bone cells, especially during bone growth or repair.     

Calycosin Suppresses RANKL-Mediated Osteoclastogenesis through Inhibition of MAPKs and NF-κB

Calycosin, an isoflavonoid phytoestrogen, isolated from Radix Astragali, was reported to possess anti-tumor, anti-inflammation, and osteogenic properties, but its impact on osteoclast differentiation remains unclear. In this study, we examined the effects of calycosin on osteoclastogenesis induced by RANKL. The results showed that calycosin significantly inhibited RANKL-induced osteoclast formation from primary bone marrow macrophages (BMMs). Calycosin also dose-dependently suppressed the formation of bone resorption pits by mature osteoclasts. In addition, the expression of osteoclatogenesis-related genes, including cathepsin K (CtsK), tartrate-resistant acid phosphatase (TRAP), and MMP-9, was significantly inhibited by calycosin. Furthermore, the results indicated that calycosin down-regulated the expression levels of NFATc1 and c-Fos through suppressing the activation of NF-κB and MAPKs. Our results indicate that calycosin has an inhibitory role in the bone loss by preventing osteoclast formation, as well as its bone resorptive activity. Therefore, calycosin may be useful as a therapeutic reagent for bone loss-associated diseases.     

Biomechanics of oral mucosa

The prevalence of prosthodontic treatment has been well recognized, and the need is continuously increasing with the ageing population. While the oral mucosa plays a critical role in the treatment outcome, the associated biomechanics is not yet fully understood. Using the literature available, this paper provides a critical review on four aspects of mucosal biomechanics, including static, dynamic, volumetric and interactive responses, which are interpreted by its elasticity, viscosity/permeability, apparent Poisson''s ratio and friction coefficient, respectively. Both empirical studies and numerical models are analysed and compared to gain anatomical and physiological insights. Furthermore, the clinical applications of such biomechanical knowledge on the mucosa are explored to address some critical concerns, including stimuli for tissue remodelling (interstitial hydrostatic pressure), pressure–pain thresholds, tissue displaceability and residual bone resorption. Through this review, the state of the art in mucosal biomechanics and their clinical implications are discussed for future research interests, including clinical applications, computational modelling, design optimization and prosthetic fabrication.     

Synthesis of absorbent polymer films made from fatty acid methyl esters,glycerol, and glutaric acid: Thermal,mechanical, and porosity analyses

In previous studies, monoglycerides (MGs) were incorporated into the matrix of poly(glutaric acid–glycerol) films to investigate their effect on the thermal, mechanical, and solvent absorption properties of the resultant films. In this study, the same properties were monitored when fatty acid methyl esters (FAME) were added to the polymer film formulation. Thermogravimetric analysis showed that, while the decomposition profile of the FAME and MG-infused films were different, the final decomposition temperatures were similar for both film types at approximately 400 °C. Degree of branching (DB%) was calculated from nuclear magnetic resonance data and was used to examine the effect of DB% on the mechanical and absorption properties of the films. Experimental results did not show any correlation with DB% and any of the physical, chemical, mechanical, or thermal properties studied. Relative to the poly(glycerol–glutaric acid) control, the incorporation of MG into the polymer matrix resulted in improved % absorption but decreased the mechanical property values. Conversely, adding FAME into the matrix improved the mechanical property values; however, there was no significant change in the % absorption values relative to the control. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47822.     

Engraftment of Prevascularized,Tissue Engineered Constructs in a Novel Rabbit Segmental Bone Defect Model

The gold standard treatment of large segmental bone defects is autologous bone transfer, which suffers from low availability and additional morbidity. Tissue engineered bone able to engraft orthotopically and a suitable animal model for pre-clinical testing are direly needed. This study aimed to evaluate engraftment of tissue-engineered bone with different prevascularization strategies in a novel segmental defect model in the rabbit humerus. Decellularized bone matrix (Tutobone) seeded with bone marrow mesenchymal stromal cells was used directly orthotopically or combined with a vessel and inserted immediately (1-step) or only after six weeks of subcutaneous “incubation” (2-step). After 12 weeks, histological and radiological assessment was performed. Variable callus formation was observed. No bone formation or remodeling of the graft through TRAP positive osteoclasts could be detected. Instead, a variable amount of necrotic tissue formed. Although necrotic area correlated significantly with amount of vessels and the 2-step strategy had significantly more vessels than the 1-step strategy, no significant reduction of necrotic area was found. In conclusion, the animal model developed here represents a highly challenging situation, for which a suitable engineered bone graft with better prevascularization, better resorbability and higher osteogenicity has yet to be developed.     

Polymorphisms of Encoding Genes IL1RN and P2RX7 in Apical Root Resorption in Patients after Orthodontic Treatment

External apical root resorption (EARR) is one of the most serious complications associated with orthodontic treatment. The aim of the study was to analyze the relationships between selected single nucleotide polymorphisms (SNPs) in Interleukin 1 receptor antagonist (IL1RN), purinoreceptor P2X7 (P2RX7) and EARR in patients after orthodontic treatment. The study comprised 101 patients who underwent a complex orthodontic treatment with a combination of fixed appliances. Roots were measured based on orthopantomograms and lateral cephalometric radiographs taken before and at the end of the treatment using diagnostic software. Proportional measurements of selected teeth were made using the modified Linge and Linge methods. Based on the presence or absence of EARR, patients were divided into two groups: control group, 61 patients without EARR (with 0.90 ≤ rRCR ≤ 1.00), and EARR group, 40 patients with EARR (rRCR < 0.90). Root resorption in selected groups was also evaluated with the scores of Malmgren and Levander. SNP analysis was performed using the real-time polymerase chain reaction (PCR) method. The analysis indicated that a specific haplotype of P2RX7 (rs208294) and IL1RN (rs419598) modified the risk of EARR development (p < 0.05), with a Bonferroni correction. The analysis of the P2RX7 and IL1RN gene polymorphisms showed that the presence of SNPs of these genes may predispose individuals to EARR. These findings indicate that EARR is a complex condition influenced not only by environmental factors and needs further study on the genetic risk factors.     

Comparison between the use of thermoplasticized gutta‐percha and a polydimethyl siloxane‐based material in filling internal resorptive cavities using spiral computed tomography

To evaluate the fill of internal resorption cavities obturated with thermoplasticized gutta‐percha and GuttaFlow2 using CT scan. Twenty human maxillary anterior teeth were selected and root canals were prepared using ProTaper system to size F3. Irrigation was performed with 5 ml of 2.5% sodium hypochlorite (NaOCl) and 5 ml of 17% ethylenediaminetetraacetic acid (EDTA). Each root was then sectioned horizontally into two halves and semicircular cavities were prepared around the periphery of the root canal opening of each root half, using a round bur. Both the root halves were then fixed using cyanoacrylate glue. All the specimens were subjected to preoperative CT scan analysis to determine the volume of internal cavities. The samples were then randomly divided into two groups. In Group 1, the specimens were obturated with thermoplasticized gutta‐percha (E&Q system) and specimens in Group 2 were obturated using GuttaFlow2. All specimens were then subjected to postoperative CT scan analysis. The volume of voids in internal resorptive cavities were calculated, which was then used to estimate the amount of gutta‐percha filled. There was no significant difference in volume of internal resorptive cavities between thermoplasticized gutta‐percha and GuttaFlow2 groups before obturation (p = 0.466). However, after obturation there was a significant difference between both the groups, in which GuttaFlow2 demonstrated better fill (p = .014). Thermoplasticized gutta‐percha filled 81% of internal resorptive cavity while GuttaFlow2 filled 91%, respectively. GuttaFlow2 showed better fill than thermoplasticized gutta‐percha in the filling of internal resorptive cavities.