Effects of vascular endothelial growth factor on osteoblasts and osteoclasts.

INTRODUCTION: Bone remodeling is crucial to the success of many dental procedures and is tightly regulated. Vascular endothelial growth factor (VEGF), a key cytokine for angiogenesis, is also an important regulator of bone remodeling. We aimed to examine the mechanisms by which VEGF induces bone remodeling by studying its effects on cultured osteoblasts and osteoclasts. METHODS: Preosteoblastic MC3T3-E1 cells were treated with vehicle or VEGF-A165. Cell proliferation, migration, and invasion potentials were assessed. Preosteoclastic RAW264.7 cells were treated with vehicle or VEGF with or without the receptor activator of nuclear factor kappa-B ligand (RANKL), and osteoclast formation was measured with tartrate-resistant acid phosphatase staining. Conditioned media from vehicle-treated or VEGF-treated MC3T3-E1 cells were tested for the levels of RANKL and osteoprotegerin (OPG) and were used to treat RAW264.7 cells to observe osteoclast formation. RESULTS: VEGF significantly induced MC3T3-E1 cell proliferation, migration, and invasion. VEGF did not directly induce osteoclastogenesis but significantly increased the RANKL/OPG ratio in the conditioned media from the MC3T3-E1 cultures; this significantly increased osteoclast formation. CONCLUSIONS: VEGF stimulates osteoclast differentiation by increasing the osteoblastic RANKL/OPG ratio but has no direct effect on osteoclast precursor cells, and it induces osteoblast proliferation, migration, and invasion potentials in vitro.

Neurologic Regulation and Orthodontic Tooth Movement.

Pain and discomfort are prevalent symptoms among the vast majority of patients with fixed orthodontic appliances and is the most disliked aspect of treatment. The periodontium is a highly innervated structure that also provides the necessary trophic factors, such as nerve growth factor, which promote neuronal survival, maintenance and axonal growth, via interaction with specific nerve surface receptors, such as TrkA. Various types of nerves are found in the periodontium, including thinly myelinated and unmyelinated sensory fibers that express the neuropeptides substance P and calcitonin gene-related peptide among others. Tooth movement activates peripheral sensory nerve endings, which transmit painful signals to the brain after being processed at the trigeminal spinal nucleus, resulting in local expression of pain related genes, such as c-Fos. Concurrently, an attendant inflammatory process is detected in the trigeminal spinal nucleus, including activation of astrocytes, microglia and neurons. This complex neurologic reaction to tooth movement mediates orthodontic pain and also serves a source of neurogenic inflammation exhibited in the trigeminal spinal nucleus and the periodontium. Activated periodontal sensory fibers release neuropeptides in the periodontal environment, which in turn induce a local inflammatory cascade aiding in alveolar bone turnover and tooth movement per se. Control of pain with nonsteroidal anti-inflammatory drugs and other prescription or over-the-counter pain killers effectively reduce this neurologic reaction and alleviate the attendant pain, but also reduce the neurogenic inflammatory component of orthodontic tooth movement causing a slowdown in bone turnover and consequently delaying orthodontic treatment.

Accelerated orthodontic tooth movement: molecular mechanisms.

Accelerating orthodontic tooth movement can significantly reduce treatment duration and risks of side effects. The rate of orthodontic tooth movement is chiefly determined by the remodeling of tissues surrounding the roots; this in turn is under the control of molecular mechanisms regulating cellular behaviors in the alveolar bone and periodontal ligament. This review summarizes the current knowledge on the molecular mechanisms underlying accelerated orthodontic tooth movement, and the clinical and experimental methods that accelerate orthodontic tooth movement with possible molecular mechanisms. The review also shows directions for future studies to develop more clinically applicable methods to accelerate orthodontic tooth movement.

Primary migration of a mini-implant under a functional orthodontic loading.

OBJECTIVES: The objective of this study was to examine whether cortical bone thickness and bone mineral density (BMD) can explain the primary migration of a mini-implant under a functional orthodontic tangential loading at the early stage following implantation. MATERIALS AND METHODS: Mini-implants were installed in human mandibular sections. A constant tangential load (2 N) was applied to the mini-implant under hydration. Creep, which is a time-dependent viscoelastic displacement in the bone surrounding the mini-implant, was assessed as the change in displacement during 2 h of loading. The total migration was measured as a maximum displacement that combined an initial elastic displacement and creep. After removal of the mini-implant, all specimens were scanned together by cone beam computed tomography. Cortical bone thickness and BMD were measured for the bone voxels surrounding the implant site. RESULTS: BMD had significant correlations with the displacement parameters (p < 0.019), but the cortical bone thickness did not (p > 0.272). Permanent bone deformation adjacent to the implant was observed to be resulting from substantial creep development under the orthodontic functional loading level. CONCLUSIONS: BMD controls the primary migration of the mini-implant system in mandibular bone. Viscoelastic creep can develop at a small constant functional loading level, leading to migration of the mini-implant. CLINICAL RELEVANCE: The current results indicated that mini-implant migration can develop under the small level of functional orthodontic load used in clinic. If the active bone remodeling around the mini-implant accelerates the migration, the risk of causing damage in vital organs next to the mini-implant increases.

Effects of orthodontic treatment on human alveolar bone density distribution.

OBJECTIVES: The objective of this study was to examine if non-invasive clinical cone beam computed tomography (CBCT)-based degree of bone mineralization (DBM) measurement can be used to detect the different results from orthodontic treatment between the maxilla and mandible in human patients. MATERIALS AND METHODS: CBCT images were taken before and after orthodontic treatment from 43 patients (19 males and 24 females, 14.36 ± 1.50 years). A histogram of computed tomography (CT) attenuation value, which is equivalent to the DBM, was obtained from the alveolar cortical (AC), trabecular (AT), and enamel (E) regions of each image. Mean, standard deviation (SD), and coefficient of variation (COV) of the CT attenuation values were computed. The regional variations and percentage (%) differences between the E and alveolar regions of the CT attenuation parameters at the maxilla and mandible were analyzed before and after orthodontic treatment. RESULTS: The AC had higher mean and variability (SD and COV) than the AT before and after treatment (p < 0.001). The variability was higher in the mandibular AC than in the maxillar AC (p < 0.01) independent of orthodontic treatment. The percentage (%) difference of variability of CT attenuation values changed for both AT and AC in the maxilla after orthodontic treatment, while that changed for only the AT (p < 0.02), but not for AC, in the mandible (p > 0.16). CONCLUSIONS: The alveolar cortical region of the mandible responded differently to orthodontic treatment compared with other alveolar regions. CLINICAL RELEVANCE: The CBCT-based DBM analysis can be used clinically to assess alveolar bone quality changes induced by orthodontic treatment to improve treatment planning and result evaluation.

Parathyroid hormone induction of cyclooxygenase-2 in murine osteoblasts: role of the calcium-calcineurin-NFAT pathway.

Murine MC3T3-E1 and MC-4 cells were stably transfected with -371/+70 bp of the murine cyclooxygenase-2 (COX-2) promoter fused to a luciferase reporter (Pluc371) or with Pluc371 carrying site-directed mutations. Mutations were made in (1) the cAMP response element (CRE) at -57/-52 bp, (2) the activating protein-1 (AP-1)-binding site at -69/-63 bp, (3) the nuclear factor of activated T-cells (NFAT)-binding site at -77/-73 bp, and (4) both the AP-1 and NFAT sites, which comprise a composite consensus sequence for NFAT/AP-1. Single mutation of CRE, AP-1, or NFAT sites decreased parathyroid hormone (PTH)-stimulated COX-2 promoter activity 40% to 60%, whereas joint mutation of NFAT and AP-1 abrogated the induction. On electrophoretic mobility shift analysis, PTH stimulated binding of phosphorylated CREB to an oligonucleotide spanning the CRE and binding of NFATc1, c-Fos, and c-Jun to an oligonucleotide spanning the NFAT/AP-1 composite site. Mutation of the NFAT site was less effective than mutation of the AP-1 site in competing binding to the composite element, suggesting that cooperative interactions of NFATc1 and AP-1 are more dependent on NFAT than on AP-1. Both PTH and forskolin, an activator of adenylyl cyclase, stimulated NFATc1 nuclear translocation. PTH- and forskolin-stimulated COX-2 promoter activity was inhibited 56% to 80% by calcium chelation or calcineurin inhibitors and 60% to 98% by protein kinase A (PKA) inhibitors. These results indicate an important role for the calcium-calcineurin-NFAT signaling pathway in the PTH induction of COX-2 and suggest that cross-talk between the cAMP/PKA pathway and the calcium-calcineurin-NFAT pathway may play a role in other functions of PTH in osteoblasts.

Anabolic effects of PTH in cyclooxygenase-2 knockout osteoblasts in vitro.

PTH is a potent bone anabolic agent in vivo but anabolic effects on osteoblast differentiation in vitro are difficult to demonstrate. This study examined the role of cyclooxygenase (COX)-2 and prostaglandin (PG) production in the effects of PTH on osteoblast differentiation in vitro using marrow stromal cell (MSC) and calvarial osteoblast (COB) cultures from COX-2 knockout (KO) and wild type (WT) mice. Cells were treated with PTH (10 nM) or vehicle throughout culture. Alkaline phosphatase (ALP) and osteocalcin (OCN) mRNA levels were measured at days 14 and 21, respectively, and mineralization at day 21. cAMP concentrations were measured in the presence of a phosphodiesterase inhibitor. PTH did not stimulate differentiation in cultures from WT mice but significantly increased ALP and OCN mRNA expression 6- to 7-fold in KO MSC cultures and 2- to 4-fold in KO COB cultures. PTH also increased mineralization in both KO MSC and COB cultures. Effects in KO cells were mimicked in WT MSC cultures treated with NS-398, an inhibitor of COX-2 activity. PTH increased cAMP concentrations similarly in WT and KO COBs. Differential gene responses to PTH in COX-2 KO COBs relative to WT COBs included greater fold-increases in the cAMP-mediated early response genes, c-fos and Nr4a2; increased IGF-1 mRNA expression; and decreased mRNA expression of MAP kinase phosphatase-1. PTH inhibited SOST mRNA expression 91% in COX-2 KO MSC cultures compared to 67% in WT cultures. We conclude that endogenous PGs inhibit the anabolic responses to PTH in vitro, possibly by desensitizing cAMP pathways.

[Flowcytometry DNA analysis of oral and maxillofacial non-Hodgkin's lymphoma].

OBJECTIVE: The purpose of this study was to investigate the relationship between the results of flowcytometry analyses of different clinical stage, location, pathologic grade and cell origin of oral and maxillofacial non-Hodgkin's lymphoma (NHL), and the diagnostic value of flowcytometry analysis in lymphoma. METHOD: This study analyzed 50 oral and maxillofacial NHL cases and 10 reactive lymph nodes (formalin fixed and paraffin embedded) by flowcytometry (FCM). RESULTS: Reactive lymph nodes were all diploid. The diploid rate of NHL was 54%, and aneuploidy rate was 46%. There was statistically significant difference between reactive lymph nodes and NHL in the DNA ploidy status and cell cycle data (SPF, CV, S + G2/M, DI). The S phase fraction (SPF) and S + G2/M had close relationship with the grade of NHL. SPF value and DNA ploidy status had no obvious relationship with the prognosis. CONCLUSION: The results suggested that the FCM had diagnostic value in NHL, especially when the morphological diagnosis was difficult. Although the cell cycle data had no prognostic value, SPF and SPF + G2/M can show the proliferative status of NHL, which can help clinical doctor select therapeutic method.