Periodontal reconstruction by heparan sulfate mimetic-based matrix therapy in Porphyromonas gingivalis-infected mice.

BACKGROUND: Periodontitis is a set of chronic inflammatory diseases affecting the supporting structures of the teeth, during which a persistent release of lytic enzymes and inflammatory mediators causes a self-perpetuating vicious cycle of tissue destruction and repair. A matrix-based therapy using a heparan sulfate (HS) analogue called ReGeneraTing Agent (RGTA) replaces destroyed HS by binding to available heparin-binding sites of structural molecules, leading to restoration of tissue homeostasis in several inflammatory tissue injuries, including a hamster periodontitis model. METHODS: The ability of RGTA to restore the periodontium was tested in a model of Porphyromonas gingivalis-infected Balb/cByJ mice. After 12 weeks of disease induction, mice were treated weekly with saline or RGTA (1.5 mg/kg) for 8 weeks. Data were analyzed by histomorphometry. RESULTS: RGTA treatment restored macroscopic bone loss. This was related to (1) a significant reduction in gingival inflammation assessed by a decrease in infiltrated connective tissue, particularly in cells expressing interleukin 1ß, an inflammatory mediator selected as a marker of inflammation; (2) a normalization of bone resorption parameters, i.e. number, activation and activity of osteoclasts, and number of preosteoclasts; (3) a powerful bone formation reaction. The Sharpey's fibers of the periodontal ligament recovered their alkaline phosphatase coating. This was obtained while P. gingivalis infection was maintained throughout the treatment period. CONCLUSIONS: RGTA treatment was able to control the chronic inflammation characteristic of periodontitis and blocked destruction of periodontal structures. It ensured tissue regeneration with recovery of the periodontium's anatomy.

NAMPT expression in osteoblasts controls osteoclast recruitment in alveolar bone remodeling.

In bone remodeling, osteoclasts are recruited via increased production of RANKL (receptor activator of nuclear factor-κB ligand) and migrate to the bone surface, aided by matrix metalloproteinases (MMPs). NAMPT (nicotinamide phosphoribosyl transferase), which catalyzes the rate-limiting step in the NAD+ salvage pathway, increases during in vitro osteogenic differentiation and inhibits RANKL-induced osteoclast differentiation. Alveolar bone loss, due to disturbance of the remodeling process, is a major feature of periodontitis. Thus, we investigated the role of NAMPT in a synchronized alveolar bone remodeling rat model. NAMPT expression increased in osteogenic cells during the remodeling activation phase, in parallel with RANKL and MMP-2 expression. Inhibition of NAMPT activity, by systemic delivery of its selective inhibitor FK866, decreased the recruitment of osteoclasts, but not their activity. In vitro, NAMPT mRNA, and protein expression also increased during osteoblast differentiation in primary calvarial osteoblast cultures. Recombinant NAMPT and NMN, its direct metabolite, dose-dependently increased bone marker expression, including that of sialoprotein (BSP) and osteocalcin (OC), whereas their expression was inhibited by FK866 treatment. Recombinant NAMPT did not regulate MMP-2, -9, MMP-13, or RANKL/OPG mRNA expression in osteoblasts. Our data suggest that de novo NAMPT synthesis in osteoblasts controls cell differentiation through osteoclast recruitment during the activation of bone remodeling.

Coordination of early cellular reactions during activation of bone resorption in the rat mandible periosteum: An immunohistochemical study.

The activation step of bone remodeling remains poorly characterized. Activation comprises determination of the site to be remodeled, osteoclast precursor recruitment, their migration to the site of remodeling, and differentiation. These actions involve different compartments and cell types. The aim of this study was to investigate events and cell types involved during activation. We used a bone remodeling model in rats where extractions of the upper jaw molars initiate remodeling of the antagonist lower jaw (mandible) cortex along the periosteum. In this model osteoclastic resorption peaks 4 days after extractions. We previously reported that mast cell activation in the periosteum fibrous compartment is an early event of activation, associated with recruitment of circulating monocyte osteoclast precursors. By using immunohistochemistry, we observed 9 hours after induction a spatially oriented expression of InterCellular Adhesion Molecule-1 in the vessels that was inhibited by antagonists of histamine receptors 1 and 2. It was followed at 12 hours by the recruitment of ED1+ monocytes. In parallel, at 9 hours, Vascular Cellular Adhesion Molecule-1+ fibroblast-like cells scattered in the fibrous compartment of the periosteum between the vessels and the osteogenic compartment increased; these cells may be implicated in osteoclast precursor migration. Receptor Activator of NF KappaB Ligand+ cells increased at 12 hours in the osteogenic compartment and reached a peak at 18 hours. At 24 hours the numbers of osteogenic cells and subjacent osteocytes expressing semaphorin 3a, a repulsive for osteoclast precursors, decreased before returning to baseline at 48 hours. These data show that during activation the two periosteum compartments and several cell types are coordinated to recruit and guide osteoclast precursors towards the bone surface.

Accelerated craniofacial bone regeneration through dense collagen gel scaffolds seeded with dental pulp stem cells.

Therapies using mesenchymal stem cell (MSC) seeded scaffolds may be applicable to various fields of regenerative medicine, including craniomaxillofacial surgery. Plastic compression of collagen scaffolds seeded with MSC has been shown to enhance the osteogenic differentiation of MSC as it increases the collagen fibrillary density. The aim of the present study was to evaluate the osteogenic effects of dense collagen gel scaffolds seeded with mesenchymal dental pulp stem cells (DPSC) on bone regeneration in a rat critical-size calvarial defect model. Two symmetrical full-thickness defects were created (5 mm diameter) and filled with either a rat DPSC-containing dense collagen gel scaffold (n = 15), or an acellular scaffold (n = 15). Animals were imaged in vivo by microcomputer tomography (Micro-CT) once a week during 5 weeks, whereas some animals were sacrificed each week for histology and histomorphometry analysis. Bone mineral density and bone micro-architectural parameters were significantly increased when DPSC-seeded scaffolds were used. Histological and histomorphometrical data also revealed significant increases in fibrous connective and mineralized tissue volume when DPSC-seeded scaffolds were used, associated with expression of type I collagen, osteoblast-associated alkaline phosphatase and osteoclastic-related tartrate-resistant acid phosphatase. Results demonstrate the potential of DPSC-loaded-dense collagen gel scaffolds to benefit of bone healing process.

Periosteum Metabolism and Nerve Fiber Positioning Depend on Interactions between Osteoblasts and Peripheral Innervation in Rat Mandible.

The sympathetic nervous system controls bone remodeling by regulating bone formation and resorption. How nerves and bone cells influence each other remains elusive. Here we modulated the content or activity of the neuropeptide Vasoactive Intestinal Peptide to investigate nerve-bone cell interplays in the mandible periosteum by assessing factors involved in nerve and bone behaviors. Young adult rats were chemically sympathectomized or treated with Vasoactive Intestinal Peptide or Vasoactive Intestinal Peptide10-28, a receptor antagonist. Sympathectomy depleted the osteogenic layer of the periosteum in neurotrophic proNerve Growth Factor and neurorepulsive semaphorin3a; sensory Calcitonin-Gene Related Peptide-positive fibers invaded this layer physiologically devoid of sensory fibers. In the periosteum non-osteogenic layer, sympathectomy activated mast cells to release mature Nerve Growth Factor while Calcitonin-Gene Related Peptide-positive fibers increased. Vasoactive Intestinal Peptide treatment reversed sympathectomy effects. Treating intact animals with Vasoactive Intestinal Peptide increased proNerve Growth Factor expression and stabilized mast cells. Vasoactive Intestinal Peptide10-28 treatment mimicked sympathectomy effects. Our data suggest that sympathetic Vasoactive Intestinal Peptide modulate the interactions between nervous fibers and bone cells by tuning expressions by osteogenic cells of factors responsible for mandible periosteum maintenance while osteogenic cells keep nervous fibers at a distance from the bone surface.

Structure and remodelling of the human parietal bone: an age and gender histomorphometric study.

OBJECTIVE: Despite its clinical usefulness, the internal structure and remodelling of parietal bone remained poorly documented. The aim of this study was to gain reliable information on parietal bone remodelling in living humans. MATERIALS AND METHODS: This study provided a site-specific analysis of static indices of turnover in relation to gender and age by using leftovers of parietal bone sampled in 100 patients (78 females; 22 males, aged 16-79 years). RESULTS: The bone architecture, cortical thickness, trabecular volume and cortical volume (C.Th, BV/TV, BV/CV) did not vary with gender. The number of osteoclasts (N.Oc/BPm) and the resorption surface (Oc.S/BS) were higher in females (p<0.05) when osteoclast resorbing activity did not vary with gender. Bone formation, osteoid surface (OS/BS) and surface covered by alkaline phosphatase-positive osteogenic cells (ALPS/BS) were higher in females (p<0.05 and p<0.01 respectively). All these parameters remained stable with aging. The osteocytic parameters, number of osteocytes (Ot.N/B.Ar) and number of osteocyte lacunae (T.L.N/B.Ar) were higher in females (p<0.05) and decreased with age in both genders (p<0.01). CONCLUSION: This study highlights the low and stable remodelling in the parietal bone. It appears to be higher in women. This stability probably reflects the low mechanical strains applied to the skull, particularly to the parietal bone.

Parietal bone as graft material for maxillary sinus floor elevation: structure and remodeling of the donor and of recipient sites.

Particulate parietal bone is used for maxillary sinus floor elevation procedure prior to dental implant placement. However, data on internal structure of the parietal bone and on graft remodeling and incorporation in the host bone are limited. We determined the structure and remodeling activities of 24 parietal bone specimens sampled at time of sinus grafting (T1 samples), and the amount and turnover of bone formed at the recipient site at time of implant placement (T2 samples, obtained 10 months after T1 samples, on average). In T1 samples, the outer cortex was 1.16+/-0.45 mm thick, had a typical haversian structure, and showed a low level of remodeling. In the cancellous portion of the samples, trabecular bone volume represented 52.8+/-10.3%. Bone remodeling was more active in the cancellous portion than in the cortical portion, but few osteoblasts and osteoclasts were seen. T2 samples consisted solely of trabecular bone, which occupied 49.4+/-18.4% of total sample volume. The boundary between new bone and the recipient bed was not discernible. Remnants of the graft particles were embedded within new bone, and showed signs of intense resorption. Bone remodeling was highly active, as shown by the presence of numerous osteoclasts resorbing new bone, together with thick osteoid seams and large osteoblasts. A loose cotton-like mineralized material was frequently observed in the marrow spaces; this acellular and non-collagenous material was strongly stained by toluidine blue, suggesting a glycoprotein nature. This study offers insights into cortical and trabecular bone structure and shows the low-level remodeling activity of parietal bone. About 10 months after grafting, the grafted chips were incorporated in new bone and almost completely resorbed. This high turnover may be beneficial for implant placement.