Murray C. Meikle

Osteogenic Gene Expression by Human Periodontal Ligament Cells under Cyclic Tension

The forces that orthodontic appliances apply to the teeth are transmitted through the periodontal ligament (PDL) to the supporting alveolar bone, leading to the deposition or resorption of bone, depending upon whether the tissues are exposed to a tensile or compressive mechanical strain. To evaluate the osteogenic potential of PDL cells, we applied a 12% uni-axial cyclic tensile strain to cultured human PDL cells and analyzed the differential expression of 78 genes implicated in osteoblast differentiation and bone metabolism by real-time RT-PCR array technology. Sixteen genes showed statistically significant changes in expression in response to alterations in their mechanical environment, including cell adhesion molecules and collagen fiber types. Genes linked to the osteoblast phenotype that were up-regulated included BMP2, BMP6, ALP, SOX9, MSX1, and VEGFA; those down-regulated included BMP4 and EGF. This study has expanded our knowledge of the transcriptional profile of PDL cells and identified several new mechanoresponsive genes.

Remodeling the Dentofacial Skeleton: The Biological Basis of Orthodontics and Dentofacial Orthopedics

Orthodontic tooth movement is dependent upon the remodeling of the periodontal ligament and alveolar bone by mechanical means. Facial sutures are also fibrous articulations, and by remodeling these joints, one can alter the positional relationships of the bones of the facial skeleton. As might be expected from the structure and mobility of the temporomandibular joint (TMJ), this articulation is more resistant to mechanical deformation, and whether functional mandibular displacement can alter the growth of the condyle remains controversial. Clinical investigations of the effects of the Andresen activator and its variants on dentofacial growth suggest that the changes are essentially dento-alveolar. However, with the popularity of active functional appliances, such as the Herbst and twin-block based on ’jumping the bite’, attention has focused on how they achieve dentofacial change. Animal experimentation enables informed decisions to be made regarding the effects of orthodontic treatment on the facial skeleton at the tissue, cellular, and molecular levels. Both rat and monkey models have been widely used, and the following conclusions can be drawn from such experimentation: (1) Facial sutures readily respond to changes in their mechanical environment; (2) anterior mandibular displacement in rat models does not increase the mitotic activity of cells within the condyle to be of clinical significance, and (3) mandibular displacement in non-human primates initiates remodeling activity within the TMJ and can alter condylar growth direction. This last conclusion may have clinical utility, particularly in an actively growing child.

Alterations in the Synthesis of IL-1β, TNF-α, IL-6, and Their Downstream Targets RANKL and OPG by Mouse Calvarial Osteoblasts In vitro: Inhibition of Bone Resorption by Cyclic Mechanical Strain

Mechanical strain is an important determinant of bone mass and architecture, and the aim of this investigation was to further understand the role of the cell–cell signaling molecules, IL-1β, TNF-α, and IL-6 in the mechanobiology of bone. Mouse calvarial osteoblasts in monolayer culture were subjected to a cyclic out-of-plane deformation of 0.69% for 6 s, every 90 s for 2–48 h, and the levels of each cytokine plus their downstream targets RANKL and OPG measured in culture supernatants by ELISAs. Mouse osteoblasts constitutively synthesized IL-1β, TNF-α, and IL-6, the production of which was significantly up-regulated in all three by cyclic mechanical strain. RANKL and OPG were also constitutively synthesized; mechanical deformation however, resulted in a down-regulation of RANKL and an up-regulation OPG synthesis. We next tested whether the immunoreactive RANKL and OPG were biologically active in an isolated osteoclast resorption pit assay – this showed that culture supernatants from mechanically deformed cells significantly inhibited osteoclast-mediated resorptive activity across the 48 h time-course. These findings are counterintuitive, because IL-1β, TNF-α, and IL-6 have well-established reputations as bone resorptive agents. Nevertheless, they are pleiotropic molecules with multiple biological activities, underlining the complexity of the biological response of osteoblasts to mechanical deformation, and the need to understand cell–cell signaling in terms of cytokine networks. It is also important to recognize that osteoblasts cultured in vitro are deprived of the mechanical stimuli to which they are exposed in vivo – in other words, the cells are in a physiological default state that in the intact skeleton leads to decreased bone strains below the critical threshold required to maintain normal bone structure.

Engineering the periodontal ligament in hyaluronan-gelatin-type I collagen constructs: upregulation of apoptosis and alterations in gene expression by cyclic compressive strain.

To engineer constructs of the periodontal ligament (PDL), human PDL cells were incorporated into a matrix of hyaluronan, gelatin, and type I collagen (COLI) in sample holders (13×1 mm) of six-well Biopress culture plates. The loading dynamics of the PDL were mimicked by applying a cyclic compressive strain of 33.4 kPa (340.6 gm/cm(2)) to the constructs for 1.0 s every 60 s, for 6, 12, and 24 h in a Flexercell FX-4000C Strain Unit. Compression significantly increased the number of nonviable cells and increased the expression of several apoptosis-related genes, including initiator and executioner caspases. Of the 15 extracellular matrix genes screened, most were upregulated at some point after 6-12 h deformation, but all were downregulated at 24 h, except for MMPs1-3 and CTGF. In culture supernatants, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinases-1 (TIMP-1) protein levels were upregulated at 24 h; receptor activator of nuclear kappa factor B (RANKL), osteoprotegerin (OPG) and fibroblast growth factor-2 (FGF-2) were unchanged; and connective tissue growth factor (CTGF) not detected. The low modulus of elasticity of the constructs was a disadvantage-future mechanobiology studies and tissue engineering applications will require constructs with much higher stiffness. Since the major structural protein of the PDL is COLI, a more rational approach would be to permeabilize preformed COLI scaffolds with PDL-populated matrices.

Appliance-induced osteopenia of dentoalveolar bone in the rat: effect of reduced bone strains on serum bone markers and the multifunctional hormone leptin

To understand, in greater detail, the molecular mechanisms regulating the complex relationship between mechanical strain and alveolar bone metabolism during orthodontic treatment, passive cross-arch palatal springs were bonded to the maxillary molars of 6-wk-old rats, which were killed after 4 and 8 d. Outcome measures included serum assays for markers of bone formation and resorption and for the multifunctional hormone leptin, and histomorphometry of the inter-radicular bone. The concentration of the bone-formation marker alkaline phosphatase (ALP) was significantly reduced at both time points in the appliance group, accompanied by a 50% reduction in inter-radicular bone volume; however, osteocalcin (bone Gla protein) levels remained unaffected. Bone collagen deoxypyridinoline (DPD) crosslinks increased 2.3-fold at 4 d only, indicating a transient increase in bone resorption; in contrast, the level of the osteoclast-specific marker, tartrate-resistant acid phosphatase 5b (TRACP 5b), was unchanged. Leptin levels closely paralleled ALP reductions at both time points, suggesting an important role in the mechanostat negative-feedback loop required to normalize bone mass. These data suggest that an orthodontic appliance, in addition to remodeling the periodontal ligament (PDL)-bone interface, may exert unexpected side-effects on the tooth-supporting alveolar bone, and highlights the importance of recognizing that bone strains can have negative, as well as positive, effects on bone mass.

Bernard G. Sarnat. 20th century plastic surgeon and biological scientist (2008)

Author: Pete E. Lestrel. Publisher: World Scientific Publishing Co. Pte Ltd, Singapore Price: £35.00 ISBN: 978-981-281-317-6 This book is a celebration of an extraordinarily long and productive life. Bernard ‘Bernie’ George Sarnat, MD, MS, DDS, FACS was born in 1912 in Chicago, the third and last child of Isadore Sarnatsky and his wife Fanny (nee Silverman). Isadore had immigrated to the USA in 1907 from Belarus following the anti-Jewish pogroms in the Russian Empire and was joined by Fanny and their two eldest children in 1909. Bernie Sarnat grew up in Chicago and was educated at Hyde Park High School and the University of Chicago (BS, 1933; MD, 1936), followed by internship at Los Angeles County General Hospital. He then decided to study dentistry, choosing the University of Illinois because of the reputation of Dr Isaac Schour, at the time the foremost biological researcher in dentistry in the USA. Illinois also had such other well-known figures as …

Northcroft Memorial Lecture 2007. A century of progress: advances in orthodontics since the foundation of the British Society for the Study of Orthodontics

The British Orthodontic Society invites outstanding contributors from the field of Orthodontics to give the guest lecture in memory of George Northcroft. In 2007 the guest lecturer was Professor Murray C. Meikle. The article that follows was presented as the Northcroft Memorial Lecture 2007 at the British Orthodontic Conference, Harrogate, UK, 24th September 2007.